Abstract Book

11th International Conference of the International Mesothelioma Interest Group Abstract Book

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FOR UPDATES DURING THE CONFERENCE! Table of Contents

NURSING PROGRAM FRIDAY SEPTEMBER 14, 2012 table of contents of table WEDNESDAY, SEPTEMBER 12, 2012 08:00-17:00 4 General Session V general information Asbestos and Mesothelioma 08:00-09:30 54

WEDNESDAY SEPTEMBER 12, 2012 Session VA1 Session IA Peritoneal Mesothelioma 10:00-11:00 56 Biomarkers for Mesothelioma Detection, Diagnosis and Prognosis 6 Session VA2 Session IB Asbestos/Nanoparticles Chemotherapy for Mesothelioma 11:00-11:30 59 10:00-11:30 10 Session VB Session IC Molecular Therapy and Genomics Gene Regulation and Mesothelioma Pathogenesis 1 10:00-11:30 60 10:00-11:30 14 Session VC Session IIA Radiology, Staging and MPM Gene Regulation and Mesothelioma Pathogenesis 2 10:00-11:30 63 14:20-16:00 18 Workshop XI Session IIB miRNA and Mesothelioma Multi-Modality 13:15-14:15 67 14:20-16:00 24 Session VIA Session IIC Novel Therapeutics: Preclinical Studies Immunology 14:20-15:50 70 14:20-16:00 27 Session VIB1 Local and Radiation Therapy THURSDAY SEPTEMBER 13, 2012 14:20-15:00 74 international mesothelioma interest group

General Session III Session VIB2 Surgical Treatment Asbestos Epidemiology 08:00-09:30 31 15:00-15:50 76

Session IIIA Session VIC1 Surgery for Mesothelioma Gene Expression Profiling 10:00-11:30 32 14:20-15:00 77

Session IIIB Session VIC2 Apoptosis and Signal Transduction Novel Therapeutics: Preclinical Studies 10:00-11:30 35 15:00-15:50 79

Session IIIC Mesothelioma Epidemiology POSTER SESSIONS | WEDNESDAY, SEPTEMBER 12, 2012 10:00-11:30 38 Poster Session 1 General Session IV Surgery New Directions and Future Studies II 11:30-12:30 82 13:15-14:15 41 Poster Session 1 Session IVA Novel Therapeutics Gene Regulation and Mesothelioma Pathogenesis 3 11:30-12:30 85 14:20-16:00 42 Poster Session 1 Session IVC Imaging and Staging Pathology 11:30-12:30 88 14:20-16:00 49 Poster Session 1 Workshop VII Pathology and Cytology Heated Chemotherapy and Other Approaches 11:30-12:30 93 to Targeting Mesothelioma Surfaces 17:00-18:30 53

iMig2012.org • Abstract Book 2 Table of Contents

Poster Session 1 POSTER SESSIONS | FRIDAY, SEPTEMBER 14, 2012 Cell Lines and Animal Models table of contents of table 11:30-12:30 96 Poster Session 5 general information Cancer Biology, Gene Regulation and Pathways and Apoptosis Poster Session 2 11:30-12:30 146 Surgery 16:00-17:00 99 Poster Session 5 Epidemiology and Asbestos Poster Session 2 11:30-12:30 153 Novel Therapeutics 16:00-17:00 104 Poster Session 5 Peritoneal Mesothelioma Poster Session 2 11:30-12:30 158 Imaging and Staging 16:00-17:00 107

Poster Session 2 AUTHOR INDEX 160 Pathology and Cytology 16:00-17:00 110

Poster Session 2 Programmatic Approach and Educational Resources for Mesothelioma Care 16:00-17:00 113

POSTER SESSIONS | THURSDAY, SEPTEMBER 13, 2012

Poster Session 3 Diagnostic and Predictive Biomarkers 11:30-12:30 116

Poster Session 3 international mesothelioma interest group Multimodality 11:30-12:30 121

Poster Session 3 Immunology 11:30-12:30 126

Poster Session 3 Radiation Therapy and Biology 11:30-12:30 128

Poster Session 4 Diagnostic and Predictive Biomarkers 16:00-17:00 130

Poster Session 4 Multimodality 16:00-17:00 135

Poster Session 4 Genomics 16:00-17:00 139

Poster Session 4 Chemotherapy and Drug Selection 16:00-17:00 142

iMig2012.org • Abstract Book 3 Nursing Program September 12, 2012 08:00-17:00

NURSING SESSION 1 GUIDING THE JOURNEY OF THE MESOTHELIOMA Low nurse to patient ratios (1:2), combined with up to date bedside P pATIENT AND FAMILY technology and monitoring tools promote nursing care interventions September 12, 2012 08:00-09:30 reflective of ‘Knowing’ the patient and family, and serve as a foundation program nusring

for establishing trust. Description of the standardized yet individualized general information NS1.2: BWH THORACIC NURSING CARING PRACTICES nursing interventions; the ‘TICU Commandments’ that are directed at UNCOVERED: LINKING CARING THEORY TO CARING PRACTICES specified patient outcomes that promote recovery will be discussed. Interdisciplinary collaboration is a cornerstone of TICU nursing care. Daily Kathleen Murphy, Carol M. Corbett Thoracic Surgery multidisciplinary rounds provide for an on unit discussion Brigham And Women’s Hospital/UNITED STATES OF AMERICA of the mesothelioma patient’s daily goals as well as post hospital care needs. Nursing representation in the weekly Thoracic Surgery Quality BWH Thoracic Surgery Conference Planning Committee Members embarked Assurance meeting allows for timely communication and ongoing care on an exploration of nursing practices used to care for the mesothelioma improvement efforts. patient and family. In addition to a comprehensive literature search, nurses reflected on patient care experiences using guided meditation, narrative Disclosure: No significant relationships. work, and story telling. Using these methods, recurrent themes emerged that illuminated the BWH specialized thoracic nursing care practices which support excellent care for mesothelioma patients and families. This nursing conference content will highlight these caring practices over each stage of NURSING SESSION 2 September 12, 2012 10:00-12:15 recovery. This talk will include discussion of identified nursing care themes, and their proposed link to the Caritas Processes described in Jean Watson’s NS2.3: THE COMPLEX DISCHARGE NEEDS OF THE MESOTHELIOMA Caring Theory. PATIENT

Disclosure: No significant relationships. Lisa Hyde-Barrett, Carol Jones-Gleeson, Kathleen Murphy, Ruben Santana Brigham And Women’s Hospital/UNITED STATES OF AMERICA

NURSING SESSION 2 September 12, 2012 10:00-12:15 Preparing the mesothelioma patient and family to leave the hospital is an integral part of the role of the thoracic nurse. Based on a two year NS2.1: SPECIALIZED NURSING CARE IN THE ICU: DELIVERING retrospective review of data the primary themes for readmission were HOPE AND POSITIVITY identified as knowledge deficits and fluid overload resulting due to a failure to follow fluid restriction. Discharge teaching is critical to prepare patients for safely transitioning home. Many factors must be taken into account, Kathryn Andrews, Paulette Puleo, Chantal Scutt, Louise Caperelli White international mesothelioma interest group Brigham And Women’s Hospital/UNITED STATES OF AMERICA including but not limited to the patient’s readiness and willingness to learn, emotional and cognitive states and cultural and educational Malignant Pleural Mesothelioma as a disease process presents many background. Understanding not only the patient’s clinical vulnerability, but challenges. Patients come to BWH for treatment with the hopes of also evaluating the patient’s emotional, psychological and social context increasing quality and quantity of life. Nurses in the Thoracic Intensive Care must be part of the plan of care. Thoracic nurses have a pivotal role in Unit are highly skilled in the care of the patient in the post op critical care the development of an optimal discharge teaching plan for mesothelioma period. The two operative procedures for the treatment of mesothelioma; patients and their caregivers. Using a case study format as a platform extrapleural pneumonectomy and pleurectomy/decortication and their for discussion, the pivotal role of the thoracic nurse during the discharge nursing care will be described. While some of the patients follow a routine process will be described. Barriers that impede patients and families from post operative course, there are others whose course is more prolonged learning how to care for themselves after discharge will be discussed and complicated. Two case studies will highlight the use of hope and and nursing strategies for overcoming those obstacles reviewed. Finally, positivity in the recovery of this patient population. Kaye Herth’s Hope the post-discharge nursing care provided to mesothelioma patients and Index and Barbara Frederickson’s Broaden and Build Theory of Positive families through an innovative nursing program created by thoracic nurses Emotion are helpful as references for our nursing practice. will be described.

Disclosure: No significant relationships.

NURSING SESSION 3 September 12, 2012 13:15-14:15

NURSING SESSION 2 September 12, 2012 10:00-12:15 NS3.1: IMP SUPPORT TEAM: CHAPLAINCY AND SOCIAL WORK

NS2.2: SPECIALIZED NURSING PRACTICES IN THE TICU: Virginia Hemani, George Winchester Sj, Charlene Haouiliya ESTABLISHING TRUST AND GUIDING RECOVERY Brigham And Women’s Hospital/UNITED STATES OF AMERICA

Teresa Breslin, Judy Finn, Rebecca Guertin, Melanie Smith, A diagnosis of Mesothelioma can be a frightening, lonely and unexpected Maureen Tapper experience; often coming as a shock to an otherwise healthy person. A Brigham And Women’s Hospital/UNITED STATES OF AMERICA cancer diagnosis affects patients, families, loved ones and friends. Its impact is not only physical, but also emotional, spiritual and financial. It is Thoracic Intermediate Care Unit (TICU) nursing care practices are directed normal to worry about how this diagnosis will affect you, your life and your towards guiding the postoperative recovery of the mesothelioma patient. family.

iMig2012.org • Abstract Book 4 The International Mesothelioma Program treats patients from all parts of the country as well as world-wide. Being far away from home adds another layer of difficulty. It means being away from supports, securing local accommodations in Boston, and all the while still coping with a life- threatening illness. To support patients and families through this process, which at the onset can feel overwhelming, there is a dedicated Mesothelioma Support Team consisting of social work, chaplaincy, a patient liaison and housing coordinator. These professionals provide counseling, emotional and spiritual support as well as practical help with housing and financial needs to patients as well as caregivers.

Disclosure: No significant relationships. nusring program nusring general information

NURSING SESSION 3 September 12, 2012 13:15-14:15

NS3.2: NURSING SUPPORT: EASING THE BURDEN OF PATIENTS AND FAMILIES

Joanne Hall, Robin Kaufman, Nancy Roy, Bernadette White Brigham And Women’s Hospital/UNITED STATES OF AMERICA

Thoracic Intermediate Care Unit nursing practice encompasses patient advocacy, establishing trust, respecting patient autonomy, and knowing the patient. These themes are central to expert Thoracic nursing practice, and have emerged through narrative work of staff nurse members of the conference planning committee. A case study presentation will explicate supportive Thoracic nursing care practices during the inpatient hospitalization of a patient with newly diagnosed mesothelioma. This skit like format coupled with the nurses reflective voice highlights notable moments in practice including patient teaching, emotional support, and specialized knowledge. The essence of the patient’s experience is highlighted and captured through practical and experienced staff familiar with the patient’s underlying pathology and resulting surgery.

Disclosure: No significant relationships.

NURSING SESSION 4 September 12, 2012 14:30-17:00

NS4.1: REFLECTIONS OF MESOTHELIOMA PATIENTS AND international mesothelioma interest group FAMILIES; IMPACT OF NURSING CARE

Meg Meccariello, Katherine Almeida Brigham And Women’s Hospital/UNITED STATES OF AMERICA

Patients and their family care givers can offer nurses valuable information about their lived experience. In this session, a Brigham & Women’s Hospital mesothelioma patient and family caregiver will speak to us about what nursing care practices were meaningful and offer suggestions on those that can be improved.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 5 Session IA Biomarkers for Mesothelioma Detection, Diagnosis and Prognosis September 12, 2012 10:00-11:30

IA.2: EARLY DETECTION OF MALIGNANT MESOTHELIOMA IN IA.3: IDENTIFICATION OF AN AUTOANTIBODY PANEL ASBESTOS-EXPOSED INDIVIDUALS WITH A NONINVASIVE AS A POTENTIAL SIGNATURE OF MALIGNANT PLEURAL PROTEOMICS-BASED SURVEILLANCE TOOL MESOTHELIOMA

Rachel M. Ostroff1, Michael Mehan1, Alex Stewart1, Deborah Ayers1, Siyu C. Zhang1, Xinbo Zhang2, Felix Fernandez-Madrid2, Harvey Pass3, Edward Brody1, Stephen Williams1, Stephen Levin2, Brad Black3, Michael Ann G. Schwartz4, Michael Harbut5 Harbut4, Michele Carbone5, Chandra Goparaju6, Harvey Pass6 1Medstart Program, Irvin D. Reid Honors College, Wayne State University, 1Somalogic, Inc., Boulder/CO/UNITED STATES OF AMERICA, 2Mt. Sinai Detroit/MI/UNITED STATES OF AMERICA, 2Internal Medicine, Wayne State Medical Center, New York/NY/UNITED STATES OF AMERICA, 3Libby University, Detroit/MI/UNITED STATES OF AMERICA, 3Cardiothoracic Mt Center For Asbestos Related Diseases/MT/UNITED STATES OF Surgery, New York University Medical Center, New York/NY/UNITED AMERICA, 4Oncology, Karmanos Cancer Institute, Detroit/MI/UNITED STATES OF AMERICA, 4Karmanos Cancer Institute, Wayne State University, STATES OF AMERICA, 5Cancer Research Center Of Hawaii/HI/UNITED Detroit/MI/UNITED STATES OF AMERICA, 5Center For Occupational And STATES OF AMERICA, 6Nyu Langone Medical Center/NY/UNITED STATES OF Environmental Medicine, Providence Hospital, Southfield/MI/UNITED AMERICA STATES OF AMERICA

Background: Malignant mesothelioma (MM) is an aggressive, asbestos- Background: Malignant pleural mesothelioma (MPM), associated with related pulmonary cancer that is increasing in incidence. Because diagnosis occupational asbestos exposure, is a deadly disease with no effective is difficult and the disease is relatively rare, most patients present at treatment due to its high resistance to chemo-radiotherapy. Molecular a clinically advanced stage where possibility of cure is minimal. To mechanisms responsible for its chemo-radiotherapeutic resistance are improve surveillance and detection of MM in the high-risk population, we complicated and undefined. The presence of side population cells (SP completed a series of clinical studies to develop a noninvasive test for early cells) in tumors is a well-accepted explanation for their anti-cancer drug detection. resistance and tumor relapse. SP cells are reported to be enriched in cancer stem cells (CSCs). Therefore, monitoring SP cells may predict both CSC- Methods: We conducted multi-center case-control studies in serum from associated aggressiveness and SP-associated chemo-radio-therapeutic 117 MM cases and 142 asbestos-exposed control individuals. Biomarker resistance. A panel of autoantibodies against SP-associated autoantigens discovery, verification, and validation were performed using SOMAmer may be an invaluable signature for early diagnosis and prediction of drug proteomic technology, which simultaneously measures over 1000 proteins resistance and MPM relapse.

in unfractionated biologic samples. international mesothelioma interest group Methods: In the present study, Hoechst 33342 SP cell analysis was used Results: Using univariate and multivariate approaches we discovered 64 to isolate SP cells from the H2714 MPM cell line. A T7 phage MPM SP cDNA protein biomarkers and derived a 13-marker random forest classifier with library was constructed in order to identify SP-associated autoantigens an AUC of 0.99 ± 0.01 in training, 0.98 ± 0.04 in independent blinded using biopan enrichment techniques with sera from patients with asbestos verification and 0.95 ± 0.04 in blinded validation studies. Sensitivity and exposure or MPM. The enrichment of SP-associated autoantigens after specificity at our pre-specified decision threshold were 97%/92% in biopanning was tested using plaque lift assay and immunochemical training and 90%/95% in blinded verification. This classifier accuracy was detection. The putative SP-associated phage clones were collected for PCR maintained in a second blinded validation set with a sensitivity/specificity and sequencing analysis. Identities of those selected autoantigens were of 90%/89% and overall accuracy of 92%. Sensitivity correlated with revealed through the sequence BLAST program. Functional annotation of pathologic stage; 77% of Stage I, 93% of Stage II, 96% of Stage III and SP-associated autoantigens was analyzed by DAVID Bioinformatics. 96% of Stage IV cases were detected. An alternative decision threshold in the validation study yielding 98% specificity would still detect 60% of Results: We have identified 300 putative SP-associated phage clones MM cases. In a paired sample set the classifier AUC of 0.99 and 91%/94% after biopan enrichment using sera of patients with asbestos exposure sensitivity/specificity was superior to that of mesothelin with an AUC of and MPM. Sequencing analysis revealed that a total of 29 clones carrying 0.82 and 66%/88% sensitivity/specificity. The biomarker panel consists of SP-associated autoantigens were in frame and unique. Table 1 shows the both inflammatory and proliferative proteins, processes strongly associated differential autoantigens identified in the sera of patients with asbestos with asbestos-induced malignancy. exposure and MPM, which summarizes BLAST results of the SP-associated autoantigen clones. Functional annotation analysis indicates that 11 Conclusion: The SOMAmer biomarker panel discovered and validated in autoantigens identified by the sera of asbestos exposed individuals are these studies provides a solid foundation for surveillance and diagnosis of related to acetylation, nucleotide binding and coupled ATPase activity, MM in those at highest risk for this disease. and that 18 autoantigens identified from the sera of MPM are involved in ribosome acetylation and WD 40 repeats, indicating that generating Disclosure: RO, MM, AS, DA, EB, and SW are employees of SomaLogic. autoantibodies against autoantigens involved in acetylation may be a common feature of asbestos-related disease. Table 1. Differential autoantigen panels identified in the sera of patients with asbestos exposure and MPM

iMig2012.org • Abstract Book 6 Results: Twenty-five MPM pts with a median follow-up of 4.2 months were included. The median age and sex ratio (M/F) were 65 years old and 2.1 respectively. Eighty-four percent of pts had an epithelioid MPM, 64% had a stage 4 disease, 60% had an anemia, a thrombocytosis or a leucocytosis at the time of inclusion. All pts except one had an Eastern Cooperative Oncology Group performance status (ECOG) < 2 and 64% received more than one line chemotherapy. CTC were detected in 48% of pts (n=12) with a median level of 1.5 (0-36). No significant correlation was observed between the presence of CTC and a metastatic disease, an ECOG ≥ 1, the presence of anaemia, leucocytosis, or thrombocytosis and the non- epithelioid type. The median PFS (1 and 2) were 17.9 (95%CI= [10.1-24.0]) and 2.5 (95%CI= [1.3-3.5]) months respectively. CTC detection was not a significant predictor of PFS 2 (p=0.27).

Conclusion: Detection of CTC has been done in a small cohort of MPM september 12, 2012 patients. Their detections could be an important tool though we were not able to demonstrate a significant prognostic value or a difference in PFS between CTC levels. Further analyzes are in progress, and updated results will be presented in September.

Disclosure: No significant relationships.

SESSION IA bIOMARKERS FOR MESOTHELIOMA DETECTION, DIAGNOSIS AND PROGNOSIS September 12, 2012 10:00-11:30 Conclusion: Differences in autoantibody signatures between the sera of patients with asbestos exposure and MPM may provide useful tool for early IA.5: THE PROGNOSTIC ROLE OF EXPRESSION OF POLO-LIKE diagnosis of MPM. Autoantibodies against SP-associated MPM autoantigens KINASE 1 (PLK1) AND CELL DIVISION CONTROL 2 (CDC2), TWO may be used to construct an invaluable panel for detecting the existence POTENTIAL THERAPEUTIC TARGETS IN MALIGNANT PLEURAL of an SP fraction within MPM, monitoring anti-cancer drug resistance, and MESOTHELIOMA predicting cancer relapse of MPM. Anthony Linton1, Kim Griggs2, Steven C. Kao1, Janette Vardy3, Stephen Disclosure: No significant relationships. Clarke4, Douglas Henderson2, Brian C. Mccaughan5, Nico Van Zandwijk1, Sonja Klebe6, Glen Reid1 1Asbestos Diseases Research Institute, Concord/NSW/AUSTRALIA, 2Flinders Medical Centre, Adelaide/AUSTRALIA, 3Concord Repatriation General SESSION IA bIOMARKERS FOR MESOTHELIOMA DETECTION, Hospital, Concord/AUSTRALIA, 4Medical Oncology, Royal North Shore DIAGNOSIS AND PROGNOSIS Hospital, St Leonards/NSW/AUSTRALIA, 5Cardiothoracic Surgical Unit, September 12, 2012 10:00-11:30 Royal Prince Alfred Hospital; The Baird Institute And Faculty Of Medicine, University Of Sydney, Newtown/AUSTRALIA, 6Department Of Anatomical IA.4: DETECTION OF CIRCULATING TUMOR CELLS IN Pathology, Filnders Univerity, Adelaide/AUSTRALIA PERIPHERAL BLOOD OF PATIENTS WITH MALIGNANT PLEURAL international mesothelioma interest group MESOTHELIOMA Background: PLK1 and CDC2 play important roles in cell cycle regulation. Elevated levels in several tumours have been associated with poor Jacques Raphael1, Christophe Massard2, Francoise Farace3, Gwénaël prognosis. Previous gene expression profiling studies revealed both targets Le Teuff4, Jacques Margery5, Fanny Billiot3, Antoine Hollebecque2, to be upregulated in malignant pleural mesothelioma (MPM). We aimed Benjamin Besse1, Jean-Charles Soria1, David Planchard1 to assess the effects of target knockdown on MPM cell growth and to 1Thoracic Group, Inserm U981, Institut Gustave Roussy, Villejuif/ determine whether the pattern of PLK1 and CDC2 expression is associated FRANCE, 2Department Of Medical Oncology, Institut Gustave Roussy, with overall survival (OS) in MPM patients. Villejuif/FRANCE, 3Translational Research Laboratory, Institut Gustave Roussy, Villejuif/FRANCE, 4Department Of Statistics And Epidemiology, Methods: Patients with a confirmed pathological diagnosis of MPM Institut Gustave Roussy, Villejuif/FRANCE, 5Pulmonary Department, Percy who underwent extrapleural pneumonectomy (EPP), or pleurectomy/ Hospital, Paris/FRANCE decortication during 1991-2009 were identified from a prospective Royal Prince Alfred Hospital database. Tissue microarrays were constructed and Background: The independent prognostic value of Circulating Tumor Cells PLK1 and CDC2 immunohistochemistry performed. OS was calculated (CTC) level has been demonstrated in patients with advanced breast, from date of diagnosis. The prognostic role of PLK1 and CDC2 was prostate and colorectal cancers. There is currently very few data on reviewed (mean score from assessable cores as percentage of tumour cells Malignant Pleural Mesothelioma (MPM) and CTC. We investigated whether labelled), adjusting for known prognostic factors including age, gender and the presence of CTC was correlated with prognosis factors and treatment histological subtype, using a Cox regression multivariate model. To assess efficacy in MPM patients. target knockdown, MPM cell lines were transfected with siRNA specific for PLK1 or CDC2, or the small molecule drugs BI 2536 and Roscovitine, and Methods: Patients (pts) with MPM in progression were enrolled before any effects on proliferation measured. new line of treatment in a prospective monocentric study. CTC detection was made on peripheral blood samples (7.5ml) using the “CellSearch” Results: 155 patients with available tissue were identified: 79% male; assay according to the manufacturer’s protocol. The correlation between median age 61 (range 22-83); 52% EPP. Histological subtypes included: the presence of CTC and known worse prognosis factors was assessed 68% epithelioid; 24% biphasic; 8% sarcomatoid. Median PLK1 expression using the X2 test. Progression Free Survival (PFS) was defined as the was 3% (Range 0-42.5%). Median CDC2 expression was 15.8% (Range time from diagnosis until first progression (PFS1) and as the time from 0.5–96%). The median OS was 14.3 months (95% CI: 10.6-18.0). On CTC measure until progression or death (PFS2). Comparison of PFS value univariate analysis, greater PLK1 expression (hazard ratio (HR) 1.90 for each according to CTC detection was performed using the log-rank test. The 10% increase; p<0.001), non-epithelioid histology (HR 3.06; p<0.001), cut-off date of the analysis was May 2012. age (HR 1.38 for each 10 year increase; p <0.001) and male gender (HR

iMig2012.org • Abstract Book 7 1.93; p = 0.004) were associated with worse outcomes. CDC2 expression pemetrexed-treated group there were no differences in overall survival was not associated. On multivariate analysis, PLK1 expression (HR 1.87; according to marker status, with two-year survival rates of 0% and 9% in 95% CI 1.23-2.83; p=0.003), subtype (HR 3.05; 95% CI 2.08-4.46; the RRM1-positive and –negative group respectively. p<0.001) and age (HR 1.22; 95% CI 1.01-1.48; p=0.036) remained significant. In mesothelioma cell lines, knockdown of either PLK1 or CDC2 Conclusion: Patients with RRM1-negative tumors treated with cisplatin- led to siRNA dose-dependent growth inhibition. Experiments with small vinorelbine combination therapy had a prolonged overall survival. There molecule inhibitors of PLK1 (BI 2536) and CDC2 (Roscovitine) also inhibited was no survival advantage in the RRM1-negative group when patients were proliferation. treated with carboplatin and pemetrexed. Our study suggest a possible role of RRM1 in predicting efficacy of cisplatin-vinorelbine combination Conclusion: We have identified PLK1 expression as an independent chemotherapy in MPM, which supports the similar finding from our group prognostic factor in MPM. Furthermore, inhibition of PLK1 and CDC2 in NSCLC patients. Thus, RRM1 may be a biomarker for vinorelbine though is growth inhibitory in MPM cells, suggesting that PLK1 and CDC2 have the results require further validation. potential as targets for therapeutic intervention in MPM. Disclosure: No significant relationships. Disclosure: No significant relationships. september 12, 2012

SESSION IA bIOMARKERS FOR MESOTHELIOMA DETECTION, SESSION IA bIOMARKERS FOR MESOTHELIOMA DETECTION, DIAGNOSIS AND PROGNOSIS DIAGNOSIS AND PROGNOSIS September 12, 2012 10:00-11:30 September 12, 2012 10:00-11:30 IA.7: BIOINFORMATICS AND MOLECULAR VALIDATION OF IA.6: AN UNEXPECTED PREDICTIVE ROLE OF RRM1 IN SEQUENTIAL BINARY GENE-RATIO TESTS IN MALIGNANT MALIGNANT PLEURAL MESOTHELIOMA TREATED WITH PLEURAL MESOTHELIOMA CISPLATIN AND VINORELBINE. Assunta De Rienzo1, William G. Richards1, Beow Y. Yeap2, Yaoyu E. Zarah G. Zimling1, Eric Santoni-Rugiu2, Cecilia Bech1, Jens B. Soerensen1 Wang3, David J. Sugarbaker1, Raphael Bueno1 1Oncology, 5074, University Hospital Of Copenhagen, Copenhagen/ 1Division Of Thoracic Surgery, Brigham And Women’s Hospital And Harvard DENMARK, 2Pathology, University Hospital Of Copenhagen, Copenhagen/ Medical School, Boston/MA/UNITED STATES OF AMERICA, 2Department DENMARK Of Medicine, Massachusetts General Hospital And Harvard Medical School, Boston/MA/UNITED STATES OF AMERICA, 3Center For Cancer Background: Ribonucleotide reductase (RNR) is an essential enzyme Computational Biology, Dana-Farber Cancer Institute, Boston/MA/UNITED for DNA synthesis that converts ribonucleoside di-phosphates into STATES OF AMERICA deoxyribonucleoside di-phosphates. The enzyme consists of a large sub-unit (RRM1) and a small sub-unit (RRM2). Overexpression of RRM1 is Background: Malignant pleural mesothelioma (MPM) is an aggressive conventionally associated with resistance towards the chemotherapeutic malignancy arising from the mesothelial cells of the pleura. Making the agent gemcitabine, which is a potent inhibitor of RNR. Our group have correct diagnosis of MPM is sometimes challenging. The epithelioid recently finished a study on the predictive value of immunohistochemically type may be difficult to distinguish from adenocarcinoma or thymoma detected RRM1 in a cohort of non small-cell lung-cancer (NSCLC) patients metastatic to the pleura, and the sarcomatoid MPM from some sarcomas from a randomized phase III trial comparing triplet-chemotherapy or other tumors with sarcomatoid histologies. We have previously showed (paclitaxel, cisplatin, gemcitabine) to standard doublet-therapy (cisplatin, that a sequential combination of binary diagnostic tests is more accurate vinorelbine). We found that increased expression of RRM1 was associated than a single diagnostic signature in diagnosing MPM from other thoracic with significantly decreased progression-free survival (PFS) and overall malignancies. We have now validated this diagnostic signature using survival (OS) only in the patients receiving cisplatin-vinorelbine therapy. bioinformatics and molecular tools. international mesothelioma interest group This finding was unexpected, since vinorelbine is a spindle-poison and resistance towards this agent has never been associated with RRM1 over- Methods: We performed expression array analyses of 112 thoracic expression. It has however been shown that vinorelbine can reduce the malignancies and control tissues using Illumina whole genome repair of radiotherapy-induced DNA damage in small-cell lung-cancer cell- microarrays. We generated 5 diagnostic tests [(calretinin/claudin-7, VACb/ lines, pointing to a possible interaction between vinorelbine and the DNA TACSTD1, MRCOX2/TITF1 for MPM vs. lung adenocarcinoma), (UBE2T/ repair system. To further investigate this finding we tested the association AGENCOURT _ 14535501; MAGED1/ADCY4; PAK4/MYH11 for MPM vs. between immunohistochemically detected RRM1 expression and outcome normal pleura), (MSLN/TGFBR3; ANXA8/TCEAL7; KRT8/PCDH18 for MPM in a cohort of patients with malignant pleural mesotheliomas (MPM) vs. sarcoma), (NFKBIZ/ARHGAP2; ARL6IP6/HPN; LOC648293/HPN for MPM treated with cisplatin and vinorelbine. vs. renal carcinoma), (RT18/PRSS16; RGS16/BCL11A, LOC648293/ PRSS16 for MPM vs. thymoma)], that were able to identify the MPM samples with Methods: Fifty-four consecutive patients with MPM, were enrolled high sensitivity and specificity. To evaluate the diagnostic power of our between February 2003 and September 2006 into a phase II trial with MPM signature using the ratio algorithm, we applied K-nearest neighbor cisplatin and vinorelbine. The formalin-fixed paraffin-embedded bioptic (KNN) and linear discrimator (LDA) analyses using Bioconductor on the tumor specimens from these MPM patients were retrospectively evaluated 26-gene signatures used to distinguish the MPM samples. Both classifiers for RRM1 expression by immunohistochemistry (IHC) using an H-score. The were trained on 70% of the randomly selected samples and tested on the cut-off point was chosen as the upper quartile value of the H-scores to remaining 30%. We then evaluated the classifiers using repeated random separate positive (H-score ≥upper quartile) from negative (H-score

iMig2012.org • Abstract Book 8 identify all the MPM samples. When we performed the sequential combination of the binary gene ratio tests using the test set we found that 90 of 100 MPM samples were correctly called as MPM, and 1 normal pleura sample and 1 sarcoma were incorrectly classified as MPM. Additional review of the MPMs called not-MPM showed that in 2 MPMs the actual specimens used for the RT-PCR had 0 % tumor content. The overall diagnostic sensitivity of the 26-gene signatures in the test set analyzed by RT-PCR was 92%, whereas the specificity was 97%.

Conclusion: Our results indicate that tests generated from a relatively a small number of genes are able to accurately distinguish MPMs from other thoracic samples supporting our hypothesis that the gene ratio tests could provide a useful clinical adjunct in the diagnosis of MPM.

Disclosure: No significant relationships. september 12, 2012 international mesothelioma interest group

iMig2012.org • Abstract Book 9 Session IB Chemotherapy for Mesothelioma September 12, 2012 10:00-11:30

SESSION IB CHEMOTHERAPY FOR MESOTHELIOMA 1Laboratory Of Molecular Biology, National Cancer Institute, Bethesda/ September 12, 2012 10:00-11:30 UNITED STATES OF AMERICA, 2Morphotek Inc./UNITED STATES OF 3

AMERICA, University Of California San Francisco, /UNITED STATES OF september 12, 2012 IB.1: UPDATED ANALYSIS OF EORTC 08983: A RANDOMIZED AMERICA, 4University Of Chicago/UNITED STATES OF AMERICA, 5University TRIAL OF RALTITREXED AND CISPLATIN VERSUS CISPLATIN Of California San Diego/UNITED STATES OF AMERICA, 6Hospital IN MALIGNANT PLEURAL MESOTHELIOMA Grosshansdorf/GERMANY, 7National Cancer Institute/UNITED STATES OF AMERICA, 8United Biosource Corporation/UNITED STATES OF AMERICA Jan P. Van Meerbeeck1, Rabab M. Gaafar2, Christian Manegold3, Sabine Margerit4, Baktiar Hasan4 Background: Amatuximab (MORAb-009) is a chimeric monoclonal 1Thoracic Oncology, Ghent University Hospital, Gent/BELGIUM, 2Medical antibody to mesothelin, a cell surface glycoprotein highly expressed in Oncology, National Cancer Institute/EGYPT, 3Thoracic Surgery, University malignant mesothelioma. Based on the results of amatuximab in a phase I Hospital/GERMANY, 4Hq, EORTC/BELGIUM clinical trial and pre-clinical studies showing synergy in combination with chemotherapy, a single arm phase II study of amatuximab plus pemetrexed Background: EORTC 08983 randomised 250 chemotherapy-naive patients (P) and cisplatin (C) was initiated in chemotherapy naïve patients with with advanced pleural mesothelioma to receive cisplatin 80 mg/m² IV on unresectable malignant pleural mesothelioma (MPM). Serum was collected day 1, alone (arm A) or combined with raltitrexed 3 mg/m² IV (arm B) (Van during the study and evaluated for potential biomarkers. Meerbeeck, J Clin Oncol 2005; 23:6881-9). Methods: Eligibility criteria included patients with unresectable epithelial Methods: An unplanned updated overall survival (OS) analysis based on or biphasic MPM, no prior chemotherapy and Karnofsky Performance the data as of May 5, 2011. OS is defined as time from randomisation to Status (KPS) >70%. A new Morphotek developed Mesothelin and MPF assay death from any cause. Adjusted indirect comparison of outcomes with was assessed as part of an exploratory end point for the clinical trial. EMPHACIS trial (Vogelzang, 2003) Results: From Feb. 2009 to Oct. 2010, 89 pts. were enrolled at 26 global Results: After a median follow up of 98 months, 118/124 participants sites. Median pt age = 67 yrs. (range 46-80), 78% male, 70% with KPS in arm A and 115/126 in arm B were deceased, an increase of 23 deaths >90%, 89% epithelial MPM, 11% biphasic MPM and 88% had stage III/ (10 and 13) compared to the previous final analysis. Seventeen (6 and 11) IV disease. In addition to the expected toxicity from PC, hypersensitivity patients were censored at the last date known to be alive: 12 (4 and 8) had reactions (12.4%; Grade 3/4=4.5%) from amatuximab were noted. Of their survival status not updated due to sites not responding, 2 in arm B the 77 evaluable pts 39% had a partial response, 51% stable disease and are alive and 3 (2 and 1) are lost to follow up. Progressive disease was the 10% progressive disease. The median PFS and OS was 6.1 months and cause of death in 87% of deaths in both arms. The median survival time 14.5 months respectively. Baseline serum mesothelin, megakaryocyte in arm B is 11.4 months (95% CI 9.5 - 13.7) compared to 8.8 months (7.7- potentiating factor (MPF) and CA-125 were assessed on all subjects.

10.7) in arm A.The hazard ratio is 0.77 (95% CI0.6 -1.0) in favour of arm B. Baseline mesothelin and MPF were found to be highly correlated with a international mesothelioma interest group The p-value for the log-rank test is 0.0491, which is still below the nominal correlation coefficient (r) =0.77, p=0.0001. Using maximally selected chi cut-off value of 0.049266 alpha level that was used at the time of the final square methodology, values of 33.14 ng/mL and 4.7 ng/mL were identified analysis. The adjusted indirect comparison of response rates, progression as optimal cut points in the determination of OS response for mesothelin and overall survival is 0.56 (95%CI 0.24-1.30), 1.15 (0.82-1.61) and 0.99 and MPF respectively. MPF was also assessed over time and its decline was (0.69-1.41), respectively. found to have modest correlation with tumor response (r=0.4; p=0.008), however no such correlation was noted with its rise and tumor progression Conclusion: The results of this updated analysis 1. confirm the superior (r =0.19; p=0.186). For CA-125, when using maximally selected chi square efficacy of the raltitrexed/cisplatin combination over cisplatin alone and 2. methodology, a value of 6 U/mL was shown to be an optimal cut point in suggest an equipoise between raltitrexed and pemetrexed in combination the determination of OS response, with the median OS of 20.7 months in with cisplatin, in the first line palliative treatment of patients with pts with baseline CA-125 less than 6 U/mL versus 13.3 months in pts with malignant pleural mesothelioma. CA-125 greater than 6 U/ml.

Disclosure: No significant relationships. Conclusion: Amatuximab in combination with P and C was generally well-tolerated with an objective response rate by independent radiological review of 39% and median OS of 14.8 months. Exploratory evaluation of potential biomarkers shows that baseline Mesothelin and MPF are highly SESSION IB CHEMOTHERAPY FOR MESOTHELIOMA correlated. In addition, baseline mesothelin, MPF and CA-125 levels were September 12, 2012 10:00-11:30 associated with improved OS and will be useful in patient selection and follow up in future studies. IB.2: UPDATED RESULTS OF THE PHASE II CLINICAL TRIAL OF THE ANTI-MESOTHELIN MONOCLONAL ANTIBODY AMATUXIMAB IN Disclosure: The following authors are employess of Morphotek Inc. D. COMBINATION WITH PEMETREXED AND CISPLATIN FOR FRONT O’Shannessy P. Fatato J. Maltzman B. Wallin J. Parno is an employee of LINE THERAPY OF PLEURAL MESOTHELIOMA AND CORRELATION United BioSource Corporation. The other authors have no disclosures. OF CLINICAL OUTCOME WITH SERUM MESOTHELIN, MPF AND CA-125

Raffit Hassan1, Dan O’Shannessy2, T. Jahan3, H. L. Kindler4, L. Bazhenova5, M. Reck6, Ira Pastan7, P Fatato2, J. Parno8, J.D. Maltzman2, B Wallin2

iMig2012.org • Abstract Book 10 SESSION IB CHEMOTHERAPY FOR MESOTHELIOMA SESSION IB CHEMOTHERAPY FOR MESTHELIOMA September 12, 2012 10:00-11:30 September 12, 2012 10:00-11:30

IB.3: RANOMIZED PHASE II TRIAL OF PEMETREXED/CISPLATIN IB.4: A PASE IB CLINICAL TRIAL OF CISPLATIN AND PEMETREXED WITH OR WITHOU CBP501 IN PATIENTS WITH ADVANCED IN COMBIATION WITH THE CD40 ACTIVATING ANTIBODY CP- MALIGNANT PLEURAL MESOTELIOMA (MPM) 870,893 IN MALINANT PLEURAL MESOTHELIOMA (MPM)

Lee M.Krug1, Antoinette Wozniak2, H.L. Kindler3, Ronald Feld4, Anna K. Nowak1, Alison Mcdonnell1, Alistair M. Cook1, Richard Lake1, Marianna Koczywas5, Jose L. Morero6, Cristina Rodriquez7, Helen Ross8, Bruce W.S. Robinson1, Jenette Creaney1, Arman Hasani2, Michael J. Julie Bauman9, Sergey Orlov10, John Ruckdeschel11, Alain Mita12, Luis Millward1 Fein13, Xiaomin He14, Robert Hall14, Takumi Kawabe15, Sunil Sharma16 1School Of Medicine And Pharmacology, University Of Western Australia, 1Department Of Medicine, Thoracic Oncology Service, Memorial Perth/WA/AUSTRALIA, 2Medical Oncology, Sir Charles Gairdner Hospital, Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF Perth/WA/AUSTRALIA AMERICA, 2Karmanos Cancer Center, Detroit/MI/UNITED STATES OF AMERICA, 3Department Of Medicine, The University Of Chicago, Chicago/ Background: CD40 is a costimulatory molecule expressed by antigen september 12, 2012 IL/UNITED STATES OF AMERICA, 4Toronto General Hospital And Princess presenting cells. Murine data supports synergy between cytotoxic Margaret Hospital, Toronto/ON/CANADA, 5City Of Hope Cancer Center, chemotherapies, including pemetrexed, and CD40 activation in a Duarte/CA/UNITED STATES OF AMERICA, 6Hospital Maria Ferrer, Buenos mesothelioma mouse model. The purpose of this study was to determine Aires/ARGENTINA, 7Oregon Health And Science Center, Portland/OR/ the maximum tolerated dose and safety profile of the CD40 activating UNITED STATES OF AMERICA, 8Mayo Clinic, Scottsdale/AZ/UNITED STATES antibody CP-870,893 in patients with advanced MPM, to explore OF AMERICA, 9University Of New Mexico, Albuquerque/NM/UNITED STATES immunological biomarkers of activity, and to provide preliminary efficacy OF AMERICA, 10St. Petersburg Medical University, St. Petersburg/RUSSIAN information. FEDERATION, 11Intermountain Healthcare, Salt Lake City/UT/UNITED STATES OF AMERICA, 12Cancer Therapy And Research Center At Uthscsa, San Methods: Eligible patients had advanced MPM, were planned for first- Antonio/TX/UNITED STATES OF AMERICA, 13Centro Oncologico De Rosario, line palliative chemotherapy with cisplatin and pemetrexed, had ECOG PS Rosario/ARGENTINA, 14Icon Clinical Research, North Wales/PA/UNITED 0-1 and adequate organ function. Patients received cisplatin (75mg/m2) STATES OF AMERICA, 15Canbas Co., Ltd., Numazu City/JAPAN, 16Huntsman and pemetrexed (500mg/m2) day 1 and CP-870,893 in escalating doses Cancer Institute, Salt Lake City/UT/UNITED STATES OF AMERICA from 0.1 mg/kg day 8 of a 21 day cycle for a maximum of 6 cycles. Stable or responding patients continued maintenance CP-870,893 D1 q21d for Background: CBP501, a synthetic duodecapeptide, is a novel compound a maximum of 6 further cycles. A standard 3+3 phase I trial design was with a dual mechanism of action: 1) CBP501 increases cisplatin influx into used. Toxicity was assessed weekly using NCI CTCAE V3.0. Immunology tumor cells through an interaction with calmodulin, enhancing cisplatin biomarkers were obtained at baseline and weekly throughout treatments. cytotoxicity, and 2) CBP501 affects cell cycle progression by abrogating CT imaging for response was performed at baseline and 6-weekly with DNA repair at the G2 checkpoint. In Phase I clinical trials of CBP501 alone measurement by Modified RECIST. or in combination with cisplatin, the most common toxicity was infusion- related urticaria managed with prophylactic antihistamines. Activity of Results: From March 2010, 15 patients received combination therapy at CBP501 plus cisplatin was observed in patients with ovarian cancer and three dose levels of CP-870,893. 3 patients were treated at dose level 1 mesothelioma, including some patients previously treated with cisplatin. (0.1mg/kg) with no dose limiting toxicities (DLT). 3 patients were treated These data prompted this randomized phase II trial in MPM. at dose level 2, with 2 DLTs (one episode of hyponatremia and confusion; one episode of splenic infarction, both occurring subsequent to cycle 1). 3 Methods: Chemotherapy naïve patients with unresectable MPM were patients were treated at dose level 1.5 (0.15 mg/kg) with no DLTs observed, stratified by histology (epithelioid vs other) and performance status (PS and an additional 6 patients were then accrued to an expansion cohort 0-1 vs 2) and randomized 2:1 to Arm A: pemetrexed/cisplatin plus CBP501 at this dose level. The best radiological response was partial response in 25 mg/m2 IV (42 pts planned), or Arm B: pemetrexed/cisplatin alone at 6 patients (40%), and stable disease in 9 patients (60%). As of May 2011, international mesothelioma interest group standard doses (21 pts planned). Patients continued on treatment until median survival is 12.9 months with 8/15 patients deceased. Two patients progression or intolerance; no maintenance single-agent therapy was are continuing single agent treatment. Two patients showed progression endorsed. The primary endpoint was progression free survival (PFS) in followed by subsequent prolonged stabilisation of disease (>24 months). Arm A; if > 23 of the 42 patients remained free of progression more than All patients experienced some grade of cytokine release syndrome (CRS) 4 months, the combination would be deemed worthy of further study. In (grade 1-2) following infusion of study drug during at least one treatment addition to standard CT imaging to assess response and PFS, PET scans, cycle. Management of CRS required patient and nursing staff education pulmonary function tests, and mesothelin levels were performed. but was readily managed using parenteral antihistamines and pethidine. Updated survival data, safety data and results of immunology studies will Results: Enrollment was completed in October 2011. 65 pts from 14 be presented. institutions were randomized, and 63 were treated. Patient characteristics in the two arms were similar in Arms A/B: median age 64/66, 80/87% Conclusion: A combination of cisplatin, pemetrexed, and CD40 activation male, 75/70% epithelioid histology. Grade 3/4 treatment-related toxicities with CP-870,893 can be safely administered in patients with advanced were uncommon, no different than expected from standard chemotherapy, MPM, and shows objective response activity similar to cisplatin and and comparable in the two arms. 53% of patients treated with CBP501 had pemetrexed alone but with a signal for a delayed and prolonged response infusion reactions, all grade 1-2. Preliminary progression free survival (PFS) in some patients. Further trials of this combination are warranted but data are available based on investigator assessments, which incorporates should include a chemotherapy-alone control arm. both radiologic and clinical progression. 27 patients (68%) in Arm A and 14 (61%) in Arm B remained free of progression greater than 4 months. The Disclosure: No significant relationships. median PFS was 5.9 mo (95% CI 4.3-9.1) in Arm A, 4.7 mo (3.7-5.9) in Arm B. The investigator-assessed best response as determined using modified RECIST in the treated population was 16/40 (40%; 95% CI 25-57) in Arm A, and 4/23 (17%; 5-39) in Arm B.

Conclusion: This randomized phase II trial met its primary endpoint. Response rate and median progression free survival favored the triplet combination arm. Updated progression data, including results from an independent radiologic review will be provided at the meeting.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 11 SESSION IB CHEMOTHERAPY FOR MESOTHELIOMA Allan Price4, Denis C. Tabot5, Tanja Cufer6, Christian Ottensmeier7, September 12, 2012 10:00-11:30 Sarah Danson8, Athanasios Pallis9, Baktiar Hasan10, Jan Van Meerbeeck11, Paul Baas12 IB.5: IFCT-GFPC-0701 MAPS TRIAL, A MULTICENTER 1Medical Oncology, Royal Marsden Hospital, /UNITED KINGDOM, 2Medical RANDOMIZED PHASE III TRIAL OF PEMETREXED-CISPLATIN WITH Oncology, National Cancer Institute, Cairo University, Cairo/ OR WITHOUT BEVACIZUMAB IN PATIENTS WITH MALIGNANT EGYPT, 3National Institute For Cancer ResearcH.L.Go Benzi, 10, / PLEURAL MESOTHELIOMA (MPM) ITALY, 4Medical Oncology, Edinburgh Cancer Research Centre/UNITED KINGDOM, 5Medical Oncology, Oxford University Hospitals Nhs Trust, Arnaud Scherpereel1, Julien Mazieres2, Jacques Margery3, Laurent European (Enets) Centre Of Excellence Oxford/UNITED KINGDOM, 6Medical Greillier4, Denis Moro-Sibilot5, Jean-jacques Parienti6, Valerie Gounant7, Oncology, University Clinic Golnik/SLOVENIA, 7Medical Oncology, Alain Riviere8, Isabelle Monnet9, Oliver Molinier10, Herve Lena11, Sylvie University Of Southampton School Of Medicine And Experimental Cancer Friard12, Jean-paul Duhamel13, Clarisse Audigier-Valette14, Gilles Medicine Centre Southampton/UNITED KINGDOM, 8Medical Oncology, Robinet15, Christian Creveuil16, Catherine Ligeza-Poisson4, Philippe Weston Park Hospital/UNITED KINGDOM, 9EORTC/BELGIUM, 10Statistics, Astoul4, Franck Morin17, Gerard Zalcman6 EORTC/BELGIUM, 11Respiratory Medicine, Gent University Hospital/ september 12, 2012 1Pulmonary And Thoracic Oncology, Lille University Hospital, Lille/ BELGIUM, 12Thoracic Oncology, Netherlands Cancer Institute/ FRANCE, 2University, Hospital Larrey, Toulouse/FRANCE, 3Oncology, NETHERLANDS Institut Gustave Roussy/FRANCE, 4Chu De Marseille/FRANCE, 5Hôpital Universitaire, Grenoble/FRANCE, 6Chu De Caen/FRANCE, 7Hopital Background: Cisplatin is one of the most active drugs available in MPM Tenon, Paris/FRANCE, 8Centre François Baclesse, Caen/FRANCE, 9Centre while bortezomib has shown some activity in single agent phase II studies Hospitalier Intercommunal De Creteil/FRANCE, 10Le Mans Regional against MPM. This was a prospective phase II study of cisplatin and Hospital, Le Mans/FRANCE, 11Chu De Rennes, Hôpital Pontchailloux, bortezomib (CB) in the first line treatment of MPM. Rennes/FRANCE, 12Hôpital Foch, Suresnes/FRANCE, 13Hôpital Prive De L’Estuaire, Le Havre/FRANCE, 14Ch Toulon/FRANCE, 15University Methods: Patients with histological proven MPM, with performance status Hospital Morvan, Brest/FRANCE, 16Centre Etienne Dolet, Saint-Nazaire/ (PS) 0/1, were eligible. The doses were cisplatin 75mg/m2 /3 wks and FRANCE, 17Ifct, Paris/FRANCE bortezomib 1.3mg/m2 day 1, 4, 8, 11 every 3 wks. The primary end-point was progression free survival rate at 18 wks (PFSR=18). The 2 stage Simon Background: MPM median OS does not exceed 13 months with design (a=0.1; b = 0.05, P0=0.50 and P1=0.675) was used. The values of P0 pemetrexed/CDDP doublet. U.S. Intergroup phase II trial of gemcitabine/ and P1 are based on results from previous EORTC LCG studies in malignant CDDP, with or without bevacizumab, gave an appealing 15.6 months mesothelioma. Data from “negative” single agent, “positive” single agent median OS in the bevacizumab arm. French Intergroup aimed to test pem/ and combination trials show that insufficient clinical activity, moderate CDDP with bevacizumab (PCB), in a randomized phase III trial. clinical activity and significant clinical activity respectively correspond to 18 week PFSR rates of 40.1%, 51.4% and 67.2%. In the first step of the Methods: Eligible patients had unresectable histologically proved MPM, study 37 eligible patients were planned. If more than 19 patients were alive no prior chemo, PS 0-2, no thrombosis, nor bleeding. Primary endpoint: and free of progression at 18 wks the total sample size was increased to 76 Primary outcome is overall survival; secondary endpoint is Progression- eligible patients. Free Survival. Patients received pem 500 mg/m2, CDDP 75 mg/m2 (PC),at D1, and vitamin B12 +B9 substitution, with (arm B) or without bevacizumab Results: Between 2007 and 2010 82 patients were entered. The median (arm A), 15 mg/kg Q21D, for 6 cycles. Arm B non- progressive patients follow-up time is 32.3 months The median age was 55 years (range: received bevacizumab maintenance therapy until progression or toxicity. 22-77yrs), male/female: 55/27 , PS 0/1: 9/73, Stage T1: 10%; T2: 42%, 445 patients to be recruited during a period 48 months, with at least 24 T3: 25%; T4: 23% and N0: 57%; N1: 4%; N2: 33%; N3: 6%. The median months of follow-up, and 385 events (deaths), will be needed to assure number of cycles received was 4 and 38% received 6 cycles. Cisplatin/ a power of 80% and detect at least a 4.3 months of median survival bortezomib dose intensity was 98/ 80%. Toxicity (grade 3/4): neutropenia increase. 10%, thrombocytopenia 11%, anaemia 1%. Grade 3-4 hyponatraemia/ hypokalaemia occurred in 46/ and 17%. Grade 2 tinnitus, grade 3 international mesothelioma interest group Results: This hypothesis leads to a Hazard Ratio (HR) of 1.33 and a 3-years fatigue occurred in 16%, and 12%, of patients. Motor/sensory/other survival of 14.7% in control arm and 23.6 % in experimental arm, with neurotoxicity was grade 1: 6/28/7%, grade 2: 2/26/2% and grade 3: an absolute difference of 8.9% in survival rates. Accrual status: The first 1/7/2% respectively. There were 2 toxic deaths at 32 and 74 days due to patient was included on February 2008. On April 30, 2012, 280 patients acute pneumonitis and cardiac arrest. The PFRS-18 (including symptomatic from 85 French centers had been enrolled. progression) was 53% (80% confidence intervals, CI, 42-64%). The overall survival was 13.5 months (95% CI 10.5-15) with 56% (95% CI 44-66%) Conclusion: The end of accrual can be expected for September 2013. alive at 1 year. The PFS was 5.1 months (95% CI 3.3-6.5). Ancillary studie: For molecular biomarker analyses, thoracoscopic tissue specimens (TS, ERCC1, MSH2, TUBB3, NF2, p16, RASSF1A methylation,15 Conclusion: On the basis of the PFRS-18, the null hypothesis could not microRNAs ) and blood samples (micro-RNAS, VEGF, osteopontin, SMRP) be rejected, CB gave predictable toxicity and was considered of moderate at diagnosis are centrally collected. Finally, a prospective study comparing activity in MPM. PET-CT to standard CT with central blinded analysis, is currently on-going for evaluation of response, and accuracy of modified RECIST criteria for Disclosure: No significant relationships. mesothelioma.

Disclosure: Research grant to AS lab and travel grant for IMIG 2012 meeting (A Scherpereel) SESSION IB CHEMOTHERAPY FOR MESOTHELIOMA September 12, 2012 10:00-11:30

IB.7: LACK OF RESPONSE TO VINORELBINE IN PATIENTS WITH SESSION IB CHEMOTHERAPY FOR MESOTHELIOMA PREVIOUSLY TREATED MALIGNANT PLEURAL MESOTHELIOMA September 12, 2012 10:00-11:30 (MPM): RATIONALE FOR PLACEBO-CONTROLLED TRIALS IN THE 2ND LINE SETTING IB.6: VALIDATION OF PFS FOR THE COMBINATION OF BORTEZOMIB (VELCADE) AND CISPLATIN AS FIRST LINE Marjorie G. Zauderer1, Camelia S. Sima2, Michelle S. Ginsberg3, TREATMENT OF MALIGNANT PLEURAL MESOTHELIOMA (MPM) Lee M. Krug1 EORTC 08052. 1Department Of Medicine, Thoracic Oncology Service, Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF Mary O’Brien1, Rabab M. Gaafar2, Sanjay K. Popat1, Francesco Grossi3, AMERICA, 2Department Of Epidemiology And Biostatistics, Memorial

iMig2012.org • Abstract Book 12 Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF AMERICA,3Department Of Radiology, Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF AMERICA

Background: After initial treatment with pemetrexed/platinum, no second- line therapy has established efficacy for MPM. Vinorelbine is listed in the NCCN guidelines as a potential treatment option for these patients based on a subgroup analysis from a first-line randomized trial (Muers, Lancet 2008) and on a single-arm, single-center phase II study (Stebbing, Lung Cancer 2009). In the latter trial, 16% of 63 patients had a partial response to vinorelbine, but these patients had not received prior treatment with pemetrexed, and over half experienced grade 3-4 toxicity. To augment the existing data, we examined our institutional experience using vinorelbine to treat patients with previously treated MPM. september 12, 2012 Methods: We reviewed the records of all patients treated at Memorial Sloan-Kettering Cancer Center with vinorelbine as second- or third- line therapy for MPM between 2003 and 2010. In all cases, vinorelbine was administered at a dose of 25 mg/m2 days 1 and 8 in a 3-week cycle. CT scans were generally performed after every two cycles. Imaging studies were reviewed according to the modified RECIST criteria.

Results: Forty-five patients were identified, including 24 treated in 2nd line and 21 in 3rd line. Patient characteristics: male 76%; median age 66 (range 41-85); epithelial 64%, sarcomatoid 19%, mixed 17%; 47% reported asbestos exposure. Treatment prior to vinorelbine included: surgery 47% (20% EPP, 27% P/D); radiation 31%; first-line therapy with pemetrexed/platinum 80%, gemcitabine/platinum 13%, and 7% other (1 patient pemetrexed alone, 2 patients on clinical trial with PDX). Table 1 summarizes the results. No complete or partial responses were achieved (95% C.I. 0-8%). Twenty patients (44%) had stable disease for a median of 2.5 months. Only 16 patients received more than 2 cycles. Seventeen patients (38%) experienced at least one episode of grade 3-4 toxicity, most commonly (>4%) neutropenia, anemia, fatigue, neutropenic fever, nausea, and vomiting.

Table 1 All(N=45) Second- Third- Line(N=24) Line(N=21) Response Rate * Complete response 0 (0%) 0 (0%) 0 (0%)

* Partial Response 0 (0%) 0 (0%) 0 (0%) international mesothelioma interest group * Stable disease 20 (44%) 9 (38%) 11 (52%) * Progression of disease 25 (56%) 15 (62%) 10 (48%) Median OS (months) 5.4 4.5 6.3 Grade 3 or 4 hematologic 12 (27%) 4 (17%) 8 (38%) toxicity

Grade 3 or 4 non- 15 (33%) 7 (29%) 8 (33%) hematologic toxicity

Conclusion: We observed no responses amongst the 45 patients in this retrospective cohort, which excludes an 8% response rate. The rate of stable disease might suggest some level of activity, and thus it remains a reasonable standard therapeutic option. However, the survival rate was comparable to that of the placebo arm in the vorinostat phase III trial (Krug, ECCO/ESMO 2011). This lack of activity supports the use of a placebo control arm in randomized second-line MPM trials.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 13 Session IC Gene Regulation and Mesothelioma Pathogenesis 1 September 12, 2012 10:00-11:30

SESSION IC GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 1 paxillin is essential for fibrillar adhesion formation. FAK, a focal adhesion September 12, 2012 10:00-11:30 kinase, preferentially interacts with paxillin and controls important

biological events, including cell migration, proliferation, and survival. We september 12, 2012 IC.1: ERK SIGNALING AND MESOTHELIOMA hypothesized that paxillin and FAK play important roles in mesothelial cell migration and thus studied the genetic aberrations, expression, and Brooke T. Mossman, Arti Shukla functionality of these molecules in mesothelioma. Pathology, University Of Vermont College Of Medicine, Burlington/VT/ UNITED STATES OF AMERICA Methods: We analyzed the expression of paxillin and FAK by IHC on 50 epithelioid, 16 sarcomatoid mesothelioma, and 1 mixed tumor tissues ERKs (Extracellular Signal- Regulated Kinases) are a family of Mitogen and compared them to 40 normal adjacent lung parenchyma. Mutational Activated Protein Kinases (MAPKs) that are linked causally to cell analysis of paxillin was performed on tumor DNA. In vitro functional proliferation and death in mesothelial and respiratory epithelial cells by analysis was performed on cells overexpressed with paxillin and its mutants asbestos and other pathogenic minerals. ERK pathways involve a cascade as well as cells with silenced paxillin. In vitro growth assays and confocal of phosphorylation events that can be stimulated by oxidative stress, imaging to define cellular motility events were performed. dimerization or activation of the EGFR, activation of B-integrins, Simian Virus 40, and chemotherapeutic drugs. Activation of ERKs culminates Results: We found an increase in paxillin and FAK expression in in induction of Activator Protein-1, a heterodimeric transcription factor mesothelioma compared to normal adjacent lung (p<0.01). Using comprised of members of the c-Fos and c-Jun proto-oncogene families automated cellular imaging system (ACIS) the immunostained slides were that governs transcription of a number of key genes in cell proliferation, quantified and the analysis of the intensity showed that normal lung transformation and malignancy. These genes and and key ERK family had IODs of 118, 13, 45 for paxillin, pFAK and FAK respectively, whereas members (ERK1, ERK2, ERK5) have been studied by our lab in mesothelial epithelioid mesothelioma had IODs of 268, 14 and 273 and sarcomatoid cells and mesotheliomas as they govern proliferation, migration, apoptosis had IODs of 331, 12 and 218. We also detected paxillin A127T mutation in and chemoresistance of MMs. Moreover, the ERK pathway is closely mesothelioma patient samples. Live-cell imaging studies revealed that in linked to the AKT tumor survival pathway and is the target of combination comparison to wild-type, mutant A127T confers a) increased lamellipodia therapies in MMs and other tumors as both ERKs and AKT levels are and filopodia formation b) enhanced mobility c) increased focal adhesion constitutively upregulated in tumor cells. Understanding the roles of formation and d) increased cell displacement in transiently transfected individual ERKs, their substrates, and cross-talk between pathways is HEK-293 cells. A mesothelial cell line with silenced paxillin showed critical to designing novel therapeutic strategies for MMs. increased sensitivity to treatment with cisplatin compared to another that did not respond similarly, emphasizing the heterogeneity of mesothelial Disclosure: No significant relationships. tumors types.

Conclusion: Paxillin and FAK are highly upregulated in sarcomatoid and international mesothelioma interest group epithelioid mesothelioma. Unique mutations were also found. Paxillin SESSION IC GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 1 mutations lead to differential processing of lamellipodia and filopodia. September 12, 2012 10:00-11:30 We believe that FAK and paxillin are important molecules in malignant mesothelioma and their biological function and therapeutic potential need IC.2: ROLE OF PAXILLIN IN MESOTHELIOMA: EXPRESSION, to be explored further. MUTATION AND FUNCTIONAL ANALYSIS Disclosure: No significant relationships. Rifat Hasina1, Ichiro Kawada1, Qudsia Arif2, Rajani Kanteti1, Maria Tretiakova2, Aliya N. Husain2, Wickii Vigneswaran3, H.L. Kindler1, Ravi Salgia1 1Medicine, The University Of Chicago, Chicago/IL/UNITED STATES OF SESSION IC GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 1 AMERICA, 2Pathology, The University Of Chicago, Chicago/IL/UNITED September 12, 2012 10:00-11:30 STATES OF AMERICA, 3Surgery, The University Of Chicago, Chicago/IL/ UNITED STATES OF AMERICA IC.3: INHIBITION OF RON (MST1R) REDUCES THE PROLIFERATIVE AND MIGRATION CAPACITY OF MESOTHELIOMA CELLS. Background: The treatment of mesothelioma has proven difficult with surgical intervention and radiation therapy due to its heterogeneic and Anne-Marie Baird1, Kenneth J. O’Byrne2, David Easty2, Alex invasive nature. Chemotherapy has been the main therapeutic option Soltermann3, Daisuke Nonaka4, Dean Fennell5, Luciano Mutti6, Harvey for many patients. More effective treatment can be achieved with Pass7, Isabelle Opitz8, Dearbhaile O’Donnell2, Steven Gray2 targeted interventions against receptor tyrosine kinases, ligands and 1Clinical Medicine, St. James’s Hospital/Trinity College Dublin, Dublin/ other intracellular proteins that modulate tumor cell growth, invasion and IRELAND, 2Clinical Medicine, Trinity College Dublin/St. James’s Hospital, metastasis. We have been studying paxillin and FAK in mesothelioma in Dublin/IRELAND, 3Division Of Clinical Pathology, University Hospital Zurich, order to analyze their role in modulating cellular mechanisms in this cancer. Zurich/SWITZERLAND, 4Department Of Pathology, New York University Paxillin is a focal adhesion phosphoprotein localized to the cytoskeleton Medical Center, New York/UNITED STATES OF AMERICA, 5Mrc Toxicology that plays a role in signal transduction, regulation of cell morphology, and Unit, University Of Leicester, Leicester/UNITED KINGDOM, 6Dept. Of the recruitment of structural and signaling molecules to focal adhesions Medicine, Vercelli Hospital, Vercelli/ITALY, 7Cardiothoracic Surgery, and is involved in motility and migration of tumor cells. It has been shown Nyu Langone Medical Center, New York/NY/UNITED STATES OF that the modulation of tyrosine phosphorylation of paxillin regulates AMERICA, 8Division Of Thoracic Surgery, University Hospital Zurich, Zurich/ both the assembly and turnover of adhesion sites and phosphorylated SWITZERLAND paxillin enhances lamellipodial protrusions whereas non-phosphorylated

iMig2012.org • Abstract Book 14 Background: RON (MST1R) is a member of the MET family and has a Methods: To study the ‘necessity’ of FGF-9 in MPM development in putative role in several cancers. The receptor has tyrosine kinase activity vivo we transfected the mouse MM cell line, AB1, with shRNA directed and consists of an alpha and a beta chain. The only ligand recognised to against murine FGF-9 (or control vector expressing a scrambled sequence). bind MST1R is the serum protein heterodimer, macrophage stimulating For the heterotopic model murine AB1-FGF-9 knock-down cells (or controls) protein (MSP). The MSP-RON signalling pathway has been implicated in were injected (5x105 cells) subcutaneously into the flank of Balb/c mice. a variety of cellular functions such as macrophage activity and wound Tumour dimensions were measured thrice weekly and animals sacrificed healing. We have previously identified MST1R/RON as frequently activated when tumours reached 100mm2 and tumour tissues harvested. FGF-9 in MPM, and high positivity for RON staining was an independent predictor expression in tumour tissue was determined by immunohistochemistry. of favourable prognosis. This study aimed to further examine the MSP-RON For orthotopic experiments, Balb/c mice received a single intraperitoneal axis in MPM. injection of 5x105 AB1-FGF-9 knock-down cells (or controls). At day 13, animals were sacrificed and the number of peritoneal tumour nodules Methods: A panel of mesothelioma cell lines were screened for the enumerated by blinded investigators. To determine whether the immune expression of MSP and MST1R at the mRNA and protein level. The system plays a role in the regulation of AB1 MM tumour growth, 5x105 AB1- proliferative response of Ju77, H226 and Met5A (non-malignant FGF-9 knock-down cells (or controls) were injected subcutaneously into transformed human pleural mesothelial cells) to recombinant MSP nude mice. To elucidate the immune cells involved in AB1 MM tumour september 12, 2012 treatment was determined. A phospho-kinase proteome profiler array was growth regulation, T cells were depleted in Balb/c tumour bearing mice utilised to detect the downstream signalling pathways activated upon MSP using specific antibodies to CD4 and CD8 and tumour growth monitored. T stimulation. Proliferation, migration and apoptotic assays were performed cell depletion was confirmed using flow cytometry. using MSP and two MST1R/RON inhibitors (a) a pre-clinical monoclonal antibody (RON8, Imclone) and (b) a small molecule inhibitor. In addition, Results: Heterotopic tumour growth was significantly retarded in mice a series of MPM TMAs were stained for MSP and macrophage (CD 68) inoculated with AB1-FGF-9 knockdown cells compared to the scrambled markers. vector and parent MM cells (p<0.001). A significant reduction in the number, and hence tumour burden, of tumour nodules was also observed Results: MSP and MST1R expression varied between the mesothelioma cell for AB1-FGF-9 knockdown tumours in the orthotopic peritoneal model line panel at both the mRNA and protein level. Treatment with recombinant compared to controls (p<0.001). When grown in nude mice, which lack MSP reduced the proliferative capacity of the Met5A cell, with a modest a functional T cell repertoire, AB1-FGF-9 knockdown tumours grew at a effect on the Ju77 MPM line. However, MSP stimulation modified the similar rate to that of the parent and vector controls which was suggestive expression of the SRC family of kinases. In terms of targeting MST1R/ of a role of the immune response in the regulation of MM tumours lacking RON, the small molecule inhibitor resulted in a significant decrease in FGF-9. AB1-FGF-9 knockdown tumours demonstrated significantly greater proliferation and migration. Although treatment with RON8 had no effect tumour burden in mice depleted of CD4+ and CD8+ T cells, either alone on proliferation, it did affect the migration capacity of the MPM cells. or in combination, when compared to saline controls which is highly High expression of MST1R/RON or MSP correlated with better survival suggestive of a T cell-mediated immune response to these tumours. by univariate analysis. In multivariate analysis, MSP was identified as an These results also suggest that FGF-9 inhibits the tumour-specific immune independent prognosticator for survival in MPM. Likewise we observed no response in MM. correlation with macrophage (CD 68) staining and survival. Conclusion: In combination with our previous in vitro data which Conclusion: The RON (MST1R) receptor comprises of a number of different clearly demonstrated the proliferative and invasive properties of FGF- isoforms, the most common of which are flRON (full length) and sf (short 9, we suggest that FGF-9 has an important role in the pathobiological form). Our study results indicate that although high levels of RON and MSP characteristics of MM in vivo and represents a novel therapeutic target. correlate with increased survival, our in vitro observations would indicate that this may be isoform dependant. Experiments are ongoing to further Disclosure: No significant relationships. elucidate the RON-MSP axis in MPM, including in vivo studies.

Disclosure: No significant relationships. international mesothelioma interest group SESSION IC GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 1 September 12, 2012 10:00-11:30

SESSION IC GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 1 IC.5: THE SURVIVIN SUPPRESSANT YM155 SELECTIVELY INHIBITS September 12, 2012 10:00-11:30 THE GROWTH OF EPITHELIOID MALIGNANT MESOTHELIOMA IN VITRO AND IN VIVO IC.4: THE ROLE OF FIBROBLAST GROWTH FACTOR-9 IN THE REGULATION OF MALIGNANT MESOTHELIOMA TUMOUR Lynette J. Schedlich, Yeun Yee Cheng, Sumedha Gattani, Ngan C. Cheng, GROWTH AND THE TUMOUR-SPECIFIC IMMUNE RESPONSE Michaela B. Kirschner, Nico Van Zandwijk, Glen Reid Asbestos Diseases Research Institute, University Of Sydney, Concord/ Sally M. Lansley1, Ai Ling Tan1, Julius Varano1, Sophia P. Karabela2, AUSTRALIA Georgios T. Stathopoulos2, Jenette Creaney3, Y C G. Lee4 1Lung Institute Of Western Australia, The University Of Western Australia, Background: Survivin plays an important role in the drug resistance of Perth/WA/AUSTRALIA, 2University Of Patras, Rio/GREECE, 3School Of many cancers. The majority of malignant pleural mesothelioma (MPM) cell Medicine And Pharmacology, National Centre For Asbestos Related lines and tumour samples express survivin, with low or no expression in Diseases, University Of Western Australia, Perth/WA/AUSTRALIA, 4Lung normal pleura. In addition, a correlation has been observed between MPM Institute Of W.A. University Of Western Australia, Perth/WA/AUSTRALIA tumours positive for survivin and shorter patient survival. YM155 is a small- molecule survivin suppressant which acts by binding to, and inactivating, Background: Identifying key molecules in the pathobiology of MM is the transcription factor ILF3, thereby reducing survivin transcription and needed to develop new therapies and biomarkers. Fibroblast growth inhibiting the growth of a broad range of human cancer cellsin vitro and in factor-9 (FGF-9) is an exciting and novel target uncovered from our vivo. This study was designed to evaluate the anti-tumour activity of global gene profiling of human MPM samples. Recently FGF-9 has been YM155 against MPM in vitro and in vivo. implicated in cancer development and neoplastic transformation of embryonic fibroblasts. We have verified over-expression of FGF-9 in MPM Methods: The effect of YM155 on survivin expression and the growth over other cancers and benign pleuritis in five separate cohorts of human and survival of MPM cells was assessed. Growth inhibitory effects pleural tissues and effusions. Our preliminary in vitro work demonstrated were measured by standard proliferation assays. Changes in cell cycle that FGF-9 induces mesothelioma cell proliferation and matrix invasion. progression and the induction of apoptosis were determined by flow We therefore hypothesised that antagonising FGF-9 may reduce tumour cytometry and caspase-3 activation, respectively. The mRNA expression aggressiveness, growth and induce tumour regression in vivo. of survivin and drug transporter genes was measured by RT-qPCR and the

iMig2012.org • Abstract Book 15 steady-state intracellular concentration of YM155 by LC-MS/MS. The anti- functions related to cell surface and nuclear syndecan-1. Experimental tumour activity of YM155, administered as a 7-day continuous infusion, settings targeting crucial cellular functions such as tumor cell proliferation, was examined in a subcutaneous MPM xenograft model. adhesion and migration have high therapeutic potential and are addressed in this project. Results: Down-regulating survivin expression in MPM cells using YM155 caused G2/M mitotic arrest, the induction of apoptosis and growth Results: Syndecan-1 overexpression had profound effects on genes inhibition. Interestingly, YM155 had greatest toxicity in cells of epithelioid involved in regulation of cell growth, cell cycle progression, adhesion, origin (IC50 range 2 – 27 nM) compared to those derived from biphasic migration and extracellular matrix organization. In particular, expression tumours (IC50 values 92 and 808 nM). YM155 decreased survivin of several growth factors, interleukins, and enzymes of importance expression at concentrations close to the IC50 value for the MPM cell line. for heparan-sulfate sulfation pattern, extracellular matrix proteins and To understand why epithelioid MPM cells are more sensitive to YM155 proteoglycans were significantly altered. 14 genes showed response to than biphasic cell lines, we investigated differences in the expression and/ both up- and down-regulation of syndecan-1. The “cytokine – cytokine- or activity of survivin and the drug transporting pumps. Survivin gene receptor interaction”, the TGF-β, EGF, VEGF and ERK/MAPK pathways were expression was lowest in the more resistant biphasic cells (R2=0.58). No enriched in both experimental settings. Most strikingly, nearly all analysed correlation was observed between the expression of uptake and efflux pathways related to cell cycle were enriched after syndecan-1 silencing and september 12, 2012 transporters and YM155 sensitivity, and their inhibition with chemical depleted after syndecan-1 overexpression. inhibitors had the same effect on YM155 sensitivity in biphasic and epithelioid cells. However, the steady-state intracellular accumulation of Conclusion: A better understanding of the complex role of syndecan-1 YM155 was 5-fold higher in epithelioid MPM cells. Interestingly, exosomes and its molecular interactions in malignant mesothelioma may provide are more abundant in media conditioned by a highly resistant biphasic cell possibilities in the future to control tumor growth and proliferation. line compared to epithelioid cells, and this may be involved in the YM155 resistance we observe. Epithelioid-selectivity was confirmed in a human Disclosure: No significant relationships. MPM xenograft model where YM155 demonstrated anti-tumour activity in epithelioid tumours but not in biphasic tumours.

Conclusion: YM155 is effective in inhibiting the growth of MPM cells, with SESSION IC GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 1 cells of epithelioid origin more sensitive than biphasic cells. The cellular September 12, 2012 10:00-11:30 basis for epithelioid selectivity is currently unknown, but may involve factors other than cellular uptake, such as increased export of drug within IC.7: NLRP3 INFLAMMASOME PLAYS A SIGNIFICANT ROLE IN exosomes. Epithelioid selectivity of YM155 has been validated in MPM THE DEVELOPMENT AND DRUG RESISTANCE OF MALIGNANT xenograft models, suggesting that YM155 may represent a promising MESOTHELIOMA subtype specific therapeutic option for MPM. Arti Shukla1, Jill M. Miller1, Maximilian B. Macpherson1, Timothy N. Disclosure: No significant relationships. Perkins1, Stacie L. Beuschel1, Harvey Pass2, Brooke T. Mossman1 1Pathology, University Of Vermont College Of Medicine, Burlington/VT/ UNITED STATES OF AMERICA, 2Cardiothoracic Surgery, Nyu Langone Medical Center, New York/UNITED STATES OF AMERICA SESSION IC GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 1 September 12, 2012 10:00-11:30 Background: Malignant mesothelioma (MM) is an aggressive cancer of mesothelial cells originating from pleural, peritoneal or pericardial IC.6: SYNDECAN-1 HAMPERS THE PROLIFERATION AND CELL- cavities. Inflammation plays an important role in development of MM. CYCLE REGULATION OF MESOTHELIOMA CELLS BY MODULATING We are first to show that asbestos activates NOD-like receptor protein 3 MULTIPLE SIGNALING PATHWAYS (NLRP3), a component of the inflammasome in human cells. As chronic asbestos exposure is a key risk factor for the development of MM, we international mesothelioma interest group Katalin Dobra1, Tünde Szatmári1, Filip Mundt2, Ghazal Heidari- hypothesized that inflammasome-mediated inflammation might underlie Hamedani1, Fang Zong1, Andrey Alexeyenko3, Anders Hjerpe1 the pathogenesis of this cancer. 1Department Of Laboratory Medicine, Karolinska Institutet, Stockholm/ SWEDEN, 2Laboratory Medicine, Karolinska Instiute, Stockholm/ Methods: To show the involvement of NLRP3 in asbestos-induced SWEDEN, 3Science For Life Laboratory/SWEDEN mesothelioma, we exposed immortalized human mesothelial cells (LP9/ hTERT), a cell type responsible for origin of MM in response to asbestos Background: Syndecan-1 is a cell surface proteoglycan (PG) important and measured steady-state NLRP3 mRNA levels by qRT-PCR, caspase-1 for the differentiation of mesothelial and epithelial cells. Dedifferentiated activation by Activity Assay kit, IL-1β release by ELISA kit and HMGB1 tumor components and mesenchymal tumours gradually loose their (High Mobility Group Box protein 1) release by Western blot analysis of cell syndecan-1 expression. In mesothelioma the expression of syndecan-1 culture supernatants. We also used human MM tumor cell lines and tumor correlates to epithelioid morphology and inhibition of growth and tissues to assess the steady-state mRNA levels of NLRP3, ASC, caspase-1 migration. Our previous data suggest a complex role of syndecan-1 in and caspase-1 activity. Small interfering RNA (siRNA) approach was used to mesothelioma cell proliferation although the exact underlying molecular depict the role of NLRP3 in asbestos-induced IL-1β and HMGB1 release. mechanisms are not completely elucidated. The aim of this study is therefore to disclose critical genes and pathways affected by syndecan-1 Results: Asbestos exposure to LP9 cells resulted in time-dependent in mesothelioma; to better understand its importance for tumour cells increases in steady-state mRNA levels and activation of NLRP3 as measured growth and proliferation. Syndecan-1 exerts its effect partly at the level of by caspase-1 activation and IL-1β release. Inhibition of NLRP3 by siRNA the cell membrane through growth factor (GFs) – growth factor receptor caused significant decreases in NLRP3 mRNA levels as well as asbestos- complexes. We have, however, shown that syndecan-1 also translocates induced IL-1β and HMGB1 release in medium. On the other hand, human to the nucleus in a regulated manner by a tubulin mediated transport MM cell lines and tumor tissues showed significantly decreased levels of mechanism. Similar nuclear transport of growth factors and their receptors NLRP3 and caspase-1 as well as caspase-1 activity as compared to LP9 or indicates a possible co-regulation with syndecan-1 and heparanase. matching normal tissues respectively.

Methods: The RMKKK motif at the cytoplasmic tail of syndecan-1 is the Conclusion: Our findings suggest that initial exposure to asbestos causes minimal sufficient sequence for this nuclear translocation. The molecular increased mRNA levels and activity of NLRP3, which may help in MM basis and function of the nuclear translocation of syndecan-1 in malignant development by promoting mesothelial cell transformation. However, mesothelioma cells are addressed by both over expression and silencing tumor development culminates in MM with decreased NLRP3 protein of syndecan-1 gene, and functional assays downstream of syndecan-1. and increased drug resistance which may in part be due to inhibition in Deletion of the RMKKK sequence allows us to separately analyze the cellular caspase-1 activity. Thus NLRP3 may be an appropriate target for therapy

iMig2012.org • Abstract Book 16 of MM, especially in combination with cytotoxic chemotherapeutic drugs and IL-1 receptor antagonists. This study is supported by a Meothelioma Applied Research Foundation (MARF) grant (AS), an NIEHS grant 1R01 ES021110-01 (AS) and by an NIEHS grants T32 ES07122 (BM).

Disclosure: No significant relationships. september 12, 2012 international mesothelioma interest group

iMig2012.org • Abstract Book 17 Session IIA Gene Regulation and Mesothelioma Pathogenesis 2 September 12, 2012 14:20-16:00

SESSION IIA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 2 SESSION IIA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 2 September 12, 2012 14:20-16:00 September 12, 2012 14:20-16:00 september 12, 2012 IIA.2: ZIC1 ACTS AS A TUMOUR SUPPRESSOR GENE AND IS IIA.3: ELUCIDATING THE RELATIONSHIP OF BAP1 TO HCF1 AND TO SILENCED IN MALIGNANT PLEURAL MESOTHELIOMA REGULATORY CHROMATIN MODIFICATIONS IN MESOTHELIOMA

Yuen Yee Cheng1, Michaela B. Kirschner1, Sonja Klebe2, J.J.B. Edelman3, Robert Mcmillan1, Matthew Bott1, Azra Krek2, Marc Ladanyi3 Michael P. Vallely3, Brian C. Mccaughan3, Rayleen V. Bowman4, Kwun 1Surgery, Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED M. Fong4, Laura Moro5, Luciano Mutti6, Hung Chuan Jin7, Nico Van STATES OF AMERICA, 2Computational Biology, Memorial Sloan-Kettering Zandwijk1, Glen Reid1 Cancer Center, New York/NY/UNITED STATES OF AMERICA, 3Pathology 1Asbestos Disease Research Institute, The University Of Sydney, Concord/ And Human Oncology & Pathogenesis Program, Memorial Sloan-Kettering AUSTRALIA, 2Department Of Anatomical Pathology, Filnders Univerity, Cancer Center, New York/NY/UNITED STATES OF AMERICA Adelaide/AUSTRALIA, 3Cardiothoracic Surgical Unit, Royal Prince Alfred Hospital; The Baird Institute And Faculty Of Medicine, University Of Sydney, Background: Frequent somatic mutations of the BAP1 nuclear Newtown/AUSTRALIA, 4Department Of Thoracic Medicine, The Prince deubiquitinase gene have recently been identified in mesothelioma (Bott Charles Hospital, Brisbane/QLD/AUSTRALIA, 5Dept. Of Pharmaceutical et al. Nat Genet 2011). Because BAP1 appears to regulate gene expression Sciences, University Of Piemonte Orientale “A. Avogadro”, Novara/ by deubiquitinating nuclear proteins such as transcription factors ITALY, 6Laboratory Of Clinical Oncology, Hospital Of Vercelli, Vercelli, Italy, (notably HCF1) and histones, we aimed to deepen our understanding of Vercelli/ITALY, 7Biomedical Research Center, Zhejiang University/CHINA BAP1’s role in gene expression by examining its genome wide location in relation to HCF1 and major histone (H) modifications, including Background: Epigenetic inactivation of tumour suppressor genes through methylation (Me), acetylation (Ac), and ubiquitination (ub), using chromatin DNA hypermethylation plays a crucial role in the progression of malignant immunoprecipitation with massively parallel sequencing (ChIP-Seq). pleural mesothelioma (MPM). ZIC1, a tumour suppressor gene silenced through promoter hypermethylation in gastric and colorectal cancer, was Methods: The BAP1-wild type MPM cell line MSTO-211H was used. To expressed at high levels in normal mesothelial cells. In contrast, ZIC1 was gauge effects on chromatin marks, experiments were performed in the not expressed or was downregulated in MPM cell lines. The aim of the presence or absence of BAP1 knockdown using BAP1 siRNA or scrambled current study was to investigate the functional significance of ZIC1 silencing siRNA, respectively. ChIP assays with antibodies to BAP1, HCF1, and the in MPM. histone marks AcH3, AcH4, H2Aub, H3K4Me, H3K9Me, H3K20Me, and H3K79Me, and H4K20Me were performed. ChIP-Seq was performed using Methods: ZIC1 mRNA expression and DNA methylation status were the Magna ChIP protocol by Millipore, and sequencing was done using the studied using RT-PCR, MSP and COBRA in MPM cell lines with and without SOLiD platform.

decitabine treatment. 24 MPM cases were included to confirm the international mesothelioma interest group mRNA expression and DNA methylation of ZIC1 using RT-PCR and MSP. Results: BAP1 was localized to approximately 1500-2500 sites along the The relationship between ZIC1 and miRNA expression was determined genome, of which only about 10% coincided with transcription start sites. by profiling miRNA expression with NCode miRNA microarrays in ZIC1- In the absence of BAP1 knockdown, approximately 25% of HCF1 sites expressing MeT-5A and MPM lines with methylation-induced ZIC1 silencing. colocalize with BAP1. BAP1 frequently colocalized with the H2Aub marks To examine the functional significance of ZIC1 silencing, ZIC1 was re- and was enriched to a lesser extent at H3K20Me marks but not at genomic expressed in MPM cell lines and effects on proliferation, migration and regions bearing the histone modifications AcH3, AcH4, H3K4Me, H3K9Me, growth in soft agar were assessed. Knockdown of ZIC1 with siRNA and and. H3K79Me. microRNA inhibitors were also included to study the functional relationship of ZIC1 silencing and microRNA expression. Conclusion: Our ChIP-Seq data so far support the known interactions of BAP1 with HCF1 and H2Aub, both of which are consistent with deregulation Results: Following treatment with decitabine, expression of ZIC1 was of gene expression as a major effect of BAP1 loss. Only a minority of BAP1 significantly up-regulated in all mesothelioma lines but was unchanged appears associated with HCF1 suggesting that many of its biological effects in MeT-5A and primary human mesothelial cells. MSP and COBRA may be through its role at H2Aub and elsewhere along the genome. More analysis revealed methylation of the ZIC1 promoter that correlated detailed analyses, including additional data, are ongoing and will be with ZIC1mRNA expression, suggesting ZIC1 is silenced in MPM through DNA presented. hypermethylation. Enforced ZIC1 expression inhibited cell migration and colony formation in H28, Ren and MM05 cell lines, and ZIC1 knockdown Disclosure: No significant relationships. enhanced growth of MeT-5A in soft agar. In MPM tumour samples ZIC1 mRNA expression was present at low or undetectable levels, with promoter methylation observed in 16 of 24 cases. Microarray analysis of MPM cell lines revealed that a number of miRNAs were overexpressed in the absence of ZIC1 expression. Upon enforced ZIC1 expression, levels of miR-23a and miR-27a were reduced, and cells transfected with an inhibitor of miR-23a exhibited reduced colony formation. These miRNAs were expressed at higher levels in tumours from patients with shorter survival.

Conclusion: Our results show that ZIC1 behaves in MPM cell culture as a tumour suppressor gene that functions in part through downregulation of miR-23a.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 18 SESSION IIA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 2 Conclusion: Somatic BAP1 mutations occur in about 20% of MPM tumors. September 12, 2012 14:20-16:00 Aside from smoking history, no other differences in clinical characteristics or outcomes were noted in the BAP1 mutated cases. Similar efforts are IIA.4: CLINICAL CHARACTERISTICS OF PATIENTS WITH needed to describe the features of germline mutations in order to define MALIGNANT PLEURAL MESOTHELIOMA (MPM) HARBORING this new cancer predisposition syndrome. We are planning a prospective SOMATIC BAP1 MUTATIONS trial to further evaluate the prevalence of germline BAP1 mutations, and we are also exploring the therapeutic implications. Marjorie G. Zauderer1, Matthew Bott2, Robert Mcmillan2, Camelia S. Sima3, Valerie Rusch4, Lee M. Krug1, Marc Ladanyi5 Disclosure: No significant relationships. 1Department Of Medicine, Thoracic Oncology Service, Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF AMERICA, 2Surgery, Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF AMERICA, 3Department Of Epidemiology And Biostatistics, Memorial Sloan-Kettering Cancer Center, New York/ NY/UNITED STATES OF AMERICA, 4Department Of Thoracic Surgery, september 12, 2012 Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF AMERICA, 5Pathology And Human Oncology & Pathogenesis Program, Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF AMERICA

Background: Efforts to elucidate tumorigenic mutations in MPM are essential to advance therapy. We reported a 23% incidence of somatic mutations in BRCA1-associated protein-1 (BAP1) in 53 patients with MPM (Bott et al. Nat Genet 2011). Germline BAP1 mutations were also reported in two families with a predisposition to mesothelioma and uveal melanoma (Testa et al. Nat Genet 2011). While BAP1 somatic mutations are more common in poor prognosis uveal melanoma (84% class 2, 4% class 1; Harbour et al. Science 2010), the significance of these mutations in MPM is unknown. Therefore, we analyzed the clinical characteristics of patients with somatic BAP1 mutations in order to describe this newly identified subpopulation.

Methods: We reviewed the charts of 121 patients with tumors tested for somatic BAP1 mutations.

Results: Patient characteristics are in Table 1. Twenty percent harbored somatic BAP1 mutations. Other than the percent of current or former smokers (75% BAP1 mutations, 42% BAP1 wild-type, p=0.006), no other clinical feature was significantly different among those with and without BAP1 mutations. Among 53 samples analyzed for NF2 mutation and p16 deletion, no correlation was seen with BAP1 mutation.

Table 1 BAP1 BAP1 p-value international mesothelioma interest group mutants wild-type (N=24) (N=97) Gender (M=male) M: 79% M: 68% 0.33 Median age at diagnosis 65 63 0.31 Histology 0.28 *Epithelial 71% 77% *Mixed 25% 12% *Sarcomatoid 4% 10% Stage 0.43 *I 13% 5% *II 25% 27% *III 33% 45% *IV 29% 22% Known asbestos exposure 54% 46% 1 Smoking (former or current) 75% 42% 0.006 Surgery 1 EPP=extrapleural pneumonectomy 54% 55% Other 38% 35% None 8% 10% Median overall survival from 14.8 15.3 0.78 diagnosis (months)

iMig2012.org • Abstract Book 19 SESSION IIA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 2 SESSION IIA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 2 September 12, 2012 14:20-16:00 September 12, 2012 14:20-16:00

IIA.5: ROLE OF HEDGEHOG SIGNALING IN MALIGNANT PLEURAL IIA.6: DEPLETION OF THE CIRCADIAN CLOCK GENE BMAL1 MESOTHELIOMA REPRESSES MALIGNANT PLEURAL MESOTHELIOMA GROWTH THROUGH INDUCTION OF MITOTIC CATASTROPHE Yangdong Shi1, Moura Ubiratan1, Isabelle Opitz2, Alex Soltermann3, Hubert Rehauer4, Svenja Thies3, Walter Weder2, Rolf Stahel1, Emanuela Momen H.K.A. Elshazley1, Mitsuo Sato1, Tetsunari Hase1, Ryo Felley-Bosco1 Yamashita1, Kenya Yoshida1, Masashi Kondo1, Futoshi Ishiguro2, Kohei 1Laboratory Of Molecular Oncology, Zurich/SWITZERLAND, 2Division Yokoi2, Noriyasu Usami2, Yoshinori Hasegawa2 Of Thoracic Surgery, University Hospital Zurich, Zurich/ 1Respiratory Medicine, Nagoya University Graduate School Of Medicine, SWITZERLAND, 3Division Of Clinical Pathology, University Hospital Zurich, Nagoya/JAPAN, 2Thoracic Surgery, Nagoya University Graduate School Of Zurich/SWITZERLAND, 4Functional Genomic Center Zurich, Zurich/ Medicine, Nagoya/JAPAN SWITZERLAND Background: Malignant pleural mesothelioma (MPM) is a highly september 12, 2012 Background: Chronic tissue inflammation and tissue repair have been aggressive neoplasm that exhibits poor prognosis, and its incidence is postulated to be the central mechanism leading to tumorigenesis in rising. Although, there has been significant progress in MPM treatment, malignant pleural mesothelioma (MPM). Tissue repair involves the development of more efficient therapeutic approaches is needed. BMAL1 is activation of stem cells and the expression of stem cell renewal genes. We a core component of the circadian clock machinery and its constitutive recently observed increased expression of PTCH1 (patched, the receptor over-expression in MPM has been reported. Here, we demonstrate binding Hedgehog ligands) in MPM side population-derived tumors which that BMAL1 serves as a molecular target to combat MPM. exhibited a tendency to have increased tumor initiating properties and developed tumors with precursor phenotype similar to tumors in patients Methods: We used 13 MPM cell lines and one non tumorgenic mesothelial with relapse after chemotherapy (Frei et al, Carcinogenesis 32: 1324, 2011). cell line (MeT-5A). In our study we performed a variety of techniques This prompted us to investigate whether HH pathway is activated in MPM including quantitative real time PCR, western blot, immuno-cytochemical,- and the effect of its inhibition in primary mesothelioma cell cultures and in histochemical and HE staining, siRNA transient transfection, growth assays a xenograft. (WST-1, liquid colony and soft agar colony formation assays), cell cycle analysis, apoptosis assay and time lapse microscopic examination. The Methods: The expression of HH signaling components was assessed clinicopathological features of 16 MPM patients were recorded for further by q-PCR and in situ hybridization in 45 clinical samples. Primary MPM analysis in correlation with BMAL1 immunohistochemical scoring in MPM cultures were developed in serum-free condition in 3% oxygen and specimens. were used to investigate the effects of Smoothened (SMO) inhibitors or GLI1 silencing on cell growth and HH signaling. In vivo effects of SMO Results: The majority of MPM cell lines and a subset of MPM antagonists were determined in a MPM xenograft growing in nude mice. clinical specimens expressed higher levels of BMAL1 compared to a non-tumorigenic mesothelial cell line (MeT-5A) and normal Results: A significant increase in GLI1, sonic hedgehog, and human parietal pleural specimens, respectively. A serum shock induced a hedgehog interacting protein gene expression was observed in MPM tumors rhythmical BMAL1expression changes in a non tumorigenic mesothelial compared to normal pleura. SMO antagonists inhibited GLI1 expression cell line, MeT-5A, but not in ACC-MESO-1 cells, suggesting that circadian and cell growth in sensitive primary cultures. This effect was mimicked rhythm pathway is deregulated in MPM cells. RNA interference-mediated by GLI1 silencing. Reduced survivin and YAP protein levels were also knockdown of BMAL1 suppressed proliferation and anchorage-dependent observed. Survivin protein levels were rescued by overexpression of GLI1 or and independent clonal growth in MPM cells but not in MeT-5A. constitutively active YAP1. Treatment of tumor-bearing mice with the Notably, BMAL1 depletion resulted in cell cycle disruption with a substantial SMO inhibitor HhAntag led to a significant inhibition of tumor growth in increase in apoptotic and polyploidy cell population in association with vivo accompanied by decreased Ki-67 and nuclear YAP immunostaining and down-regulation of Wee1, cyclin B, and p21WAF1/CIP1 and up-regulation of international mesothelioma interest group a significant difference in selected gene expression profile in tumors. cyclin E expression. ACC MESO-1 cells exhibited drastic morphological changes including micronucleation, multiple nuclei and increased cellular Conclusion: An aberrant HH signaling is present in MPM and inhibition of volume and time lapse microscopic examination demonstrates mitotic HH signaling decreases tumor growth indicating potential new therapeutic catastrophe as a cell fate following BMAL1-knockdown in ACC MESO-1 cells approach. that expressed the highest level of BMAL1.

Disclosure: No significant relationships. Conclusion: In conclusion, we provide evidence that BMAL1 has a critical role in MPM and could serve as an attractive therapeutic target for MPM.

Disclosure: No significant relationships.

(see graphic next page)

iMig2012.org • Abstract Book 20 september 12, 2012 international mesothelioma interest group

iMig2012.org • Abstract Book 21 SESSION IIA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 2 SESSION IIA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 2 September 12, 2012 14:20-16:00 September 12, 2012 14:20-16:00

IIA.7: CAN HDAC/ER BETA EXPRESSION BE USED TO STRATIFY IIA.8: THE BAP1 CANCER SYNDROME MESOTHELIOMA PATIENTS FOR APPROPRIATE TREATMENT 1 2 1 REGIMENS Michele Carbone , Laura Korb Ferris , Francine Baumann , Andrea Napolitano1, Christopher A. Lum3, Erin G. Flores1, Giovanni Gaudino1, 1 4 5 Anne-Marie Baird1, Louise Flynn2, Eimear O’Donnell2, Cormac J. Amy Powers , Peter Bryant-Greenwood , Thomas Krausz , Elizabeth 5 6 7 8 9 Jennings3, Martin P. Barr2, Eric Santoni-Rugiu4, Jens Benn Sørensen5, Hyjek , Rachael Tate , Joseph Friedberg , Tracey Weigel , Harvey Pass , 10 Zarah G. Zimling5, Warren Thomas3, Luciano Mutti6, Laura Moro7, Haining Yang 1 Kenneth J. O’Byrne2, Steven Gray2 University Of Hawaii Cancer Center, Honolulu/UNITED STATES 2 1Clinical Medicine, St. James’s Hospital/Trinity College Dublin, Dublin/ OF AMERICA, Dermatology, University Of Pittsburgh, Pittsburgh/ 3 IRELAND, 2Clinical Medicine, Trinity College Dublin/St. James’s Hospital, UNITED STATES OF AMERICA, Molecular Diagnostics And Dublin/IRELAND, 3Department Of Molecular Medicine, Royal College Of Biorepository, Queen’s Medical Center, Honolulu/UNITED STATES OF

4 september 12, 2012 Surgeons In Ireland, Dublin/IRELAND, 4Pathology, University Hospital AMERICA, Pathology, Queen’s Medical Center, Honolulu/UNITED STATES 5 Of Copenhagen, Copenhagen/DENMARK, 5Oncology, 5074, University OF AMERICA, Pathology, University Of Chicago, Chicago/UNITED 6 Hospital Of Copenhagen, Copenhagen/DENMARK, 6Dept. Of Medicine, STATES OF AMERICA, Private Practice, Opelousas/UNITED STATES OF 7 Vercelli Hospital, Vercelli/ITALY, 7Dept. Of Pharmaceutical Sciences, AMERICA, Thoracic Surgery, Unveristy Of Pennsylvania, Philadelphia, 8 University Of Piemonte Orientale “A. Avogadro”, Novara/ITALY PA/UNITED STATES OF AMERICA, Surgery, University Of Wisconsin, Madison, WI/UNITED STATES OF AMERICA, 9Cardiothoracic Surgery, Background: Malignant pleural mesothelioma (MPM) is a rare and New York University Medical Center, New York/NY/UNITED STATES OF aggressive cancer. The severity of this disease is underscored by the fact AMERICA, 10University Of Hawaii Cancer Center, Honolulu, Hi/UNITED that no single treatment option has proven particularly effective. The STATES OF AMERICA current standard of care for patients suffering from MPM is a combination of pemetrexed and cisplatin with an observed objective response rate Background: In 1997 the extremely high incidence of mesothelioma in of approximately 40%. Accordingly there is an urgent need to identify certain Cappadocian families suggested to me that genetics was playing a patients who may benefit from this regimen, and furthermore identify new role. We discovered that mesothelioma was transmitted in an autosomal translational targets or approaches that potentially could be used to treat dominant fashion and we started the hunt for a putative mesothelioma MPM patients. Epigenetic modifiers such as histone deacetylases (HDACs) gene (Roushdy-Hammady I., et al., The Lancet, 2001; Dogan U., et expression levels may be useful to stratify patients into treatment regimens al., Cancer Res 2006; Carbone et al., Nat Rev Cancer 2007). Environmental such as those that will respond to (a) HDAC inhibition, (b) Estrogen and mineralogical studies revealed that Cappadocian families with high Receptor targeted agents and (c) those cisplatin therapy. and low incidence of mesothelioma were exposed to similar amounts of erionite (Carbone M., et al., PNAS 2011). We discovered that germline Methods: Fifteen mesothelioma and one normal cell line (Met5A) were BAP1 mutations are associated with high incidence of mesothelioma and screened for the expression of HDAC11, Class I (HDAC1, 2, 3, and 8) and uveal melanoma (UVM), and that BAP1 is frequently mutated in sporadic Class II (HDAC4, 5, 6, 7, 9, 10) histone deacetylases at (a) the protein level mesothelioma (Testa JR et al., Nat Genet 2011). Speicher’s team discovered by means of Western Blot and (b) the mRNA level using RT-PCR. The HDAC that germline BAP1 mutations caused benign melanocytic tumors (Wiesner expression profile of a cisplatin resistant cell line (P31) was also determined. T., et al, (Nat Genet 2011). Additionally HDAC (HDAC1, 2, 3) and ER-Beta expression was examined in panel of twenty patient samples (benign, biphasic, sarcomatoid, Methods: We investigated for melanocytic lesions the US families with epithelial). high incidence of mesothelioma and we conducted a meta-analysis of the published studies of families carrying germline BAP1 mutations. Results: HDAC11 and Class I and II HDACs were detected to varying degrees within the mesothelioma and normal cell lines. HDAC1/2 and 4 were Results: We discovered that germline BAP1 mutations cause a new international mesothelioma interest group universally expressed at the protein level with HDAC3 (8/16), HDAC7/8 cancer syndrome characterized by mesothelioma, uveal and cutaneous (13/16), HDAC5 (14/16) demonstrating differential expression. In the P31 melanoma, MBAITs (melanocytic BAP1-mutated atypical intradermal cell line, HDAC protein expression was decreased (HDAC2/3/4/5/7) in the tumors”), and possibly renal cell and other carcinomas (Carbone M., cisplatin resistant sub type compared with the parent. Furthermore HDAC5, et al., JTM in press). MBAITs are benign melanocytic tumors that have was significantly reduced (p<0.05) in the cisplatin resistant cell line. distinct histological and molecular characteristics when compared to Presently a cohort of mesothelioma patient samples is undergoing IHC other melanocytic lesions. MBAITs develop early in life and allow the staining for HDAC5. Expression of HDAC1/2 and 3 were increased in the identification of potential carriers of germline BAP1 mutations who can MPM patient samples (n=15) compared with benign (n=5) (HDAC2/3, be followed for early detection of the malignancies associated with this p<0.05). Overall, ER-Beta protein levels were decreased in the MPM syndrome. samples compared with benign. An apparent inverse correlation between ER-Beta and HDAC expression was observed. Conclusions: The capacity of BAP1 mutations to cause multiple tumor types and the very high tumor phenotype penetrance (close to 100% in these Conclusion: This is one of the first studies to determine HDAC expression families) indicates that this gene plays a major role in influencing cancer in clinically relevant patient samples. Altered HDAC expression was cell growth. The pleiotropic effects of BAP1 can account for this finding. observed in an isogenic parent and cisplatin resistant cell model, which We advise families with hereditary BAP1 mutation to have family members may suggest that a reduction in HDAC expression is involved in cisplatin tested for mutant gene carrier status at the age ten and, if positive, to resistance in MPM. Conversely, an increase in the protein levels of HDAC1/2 begin routine screening with a total body dermatological examination and and 3 were detected in MPM patient samples with decreased levels of annual eye examinations as family members may develop melanoma at an ER-Beta. An inverse correlation was evident between ER-Beta and HDAC early age. expression within our patient cohort. This suggests that HDAC expression could be used to stratify patients for treatment regimens.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 22 september 12, 2012

Cancers associated to BAP1 syndrome: dimension of circles is proportional to cancer prevalence in BAP1-mutated families. Solid arrows indicate the tumors associated to BAP1 syndrome. Dashed arrows indicate cancers possibly associated to the syndrome. CM: cutaneous melanoma; MM: malignant mesothelioma; CCRC: clear cell renal carcinoma

Disclosure: No significant relationships. international mesothelioma interest group

iMig2012.org • Abstract Book 23 Session IIB Multi-Modality September 12, 2012 14:20-16:00

SESSION IIB mULTI-MODALITY SESSION IIB mULTI-MODALITY September 12, 2012 14:20-16:00 September 12, 2012 14:20-16:00 september 12, 2012 IIB.2: LONG-TERM OUTCOME AFTER RADICAL PLEURECTOMY IIB.3: PERCUTANEOUS OUTPATIENT CRYOABLATION FOR FOLLOWED BY CHEMORADIATION FOR MALIGNANT PLEURAL LOCALIZED RECURRENT PLEURAL MESOTHELIOMA FOLLOWING MESOTHELIOMA: A 10-YEAR SINGLE CENTER EXPERIENCE LUNG-SPARING PLEURECTOMY AND DECORTICATION SURGERY

Servet Bölükbas1, Michael Eberlein2, Natalie Kudelin1, Annette Fisseler- Fereidoun Abtin1, Robert Suh1, Anne Rorie2, Jesse Sandberg3, Robert B. Eckhoff3, Joachim Schirren1 Cameron2 1Thoracic Surgery, Dr. Horst Schmidt Klinik, Wiesbaden/GERMANY, 2Division 1Radiology, David Geffen School Of Medicine At UCLA, Los Angeles/CA/ Of Pulmonary, Critical Care And Occupational Medicine, Carver UNITED STATES OF AMERICA, 2Thoracic Surgery, David Geffen School Of College Of Medicine, University Of Iowa, Iowa/UNITED STATES OF Medicine At UCLA, Los Angeles/CA/UNITED STATES OF AMERICA, 3David AMERICA, 3Institutes For Pathology And Cytology, Dr. Horst Schmidt Klinik, Geffen School Of Medicine At UCLA, Los Angeles/CA/UNITED STATES OF Wiesbaden/GERMANY AMERICA

Background: We report our 10-year single center experience of malignant Background: Patients with malignant pleural mesothelioma have a high pleural mesothelioma (MPM) treated with radical pleurectomy (RP) as local recurrence rate following surgery with or without postoperative surgical arm within a trimodality approach. adjuvant therapy. Once recurrent, control of the disease is difficult, particularly while maintaining a satisfactory quality of life. We sought to Methods: In a prospective, non-randomized study, all patients with assess the efficacy and quality of life impact of percutaneous CT-guided histologically proven MPM, clinical stage cT1-3 cN0-2 and without prior cryoablation in the control of localized recurrence following pleurectomy treatment for MPM were evaluated for trimodality therapy: lung-sparing RP and decortication for pleural mesothelioma. followed by 4 cycles of chemotherapy (Cisplatin/Pemetrexed) and radiation of the chest wall at the intervention sites from 2002 to 2011. Kaplan-Meier Methods: Following institutional IRB approval, we retrospectively reviewed analyses, log–rank test and Cox regression analyses were used to estimate our prospective thoracic surgery and radiology databases for patients who survival and to determine predictors of survival. were identified as having one or more cryoablation treatments for localized recurrence of malignant pleural mesothelioma following surgery with or Results: Eighty-eight out of 206 consecutive patients underwent RP without adjuvant therapy. followed by chemoradiation. 74 out of 88 patients (84%) completed the trimodality therapy. Surgical morbidity and mortality were 27.3% (24/88) Results: We identified 24 patients who had undergone one or more and 2.3% (2/88), respectively. Median survival (MS) was 26 months (mo). cryoablation procedures for localized recurrent malignant pleural

One-, 3- and 5-year-survival were 78%, 30% and 25%, respectively. mesothelioma. Mean patient age was 63.5 years. 13/24 (54.2%) male international mesothelioma interest group Progression-free-survival was 13 Mo. The sites of failure were locoregional and 11/24 (45.8%) female. Histologies were epithelioid predominant in (47.6%), distant (11.0%) and both (13.4%). Median time between disease 19/24 (79.2%), mixed in 4/24 (16.6%), and sarcomatoid in 1/24 (4.2%). progression and death was 7 Mo. Incomplete resections (p<0.001), All patients underwent surgery and 22/24 patients (91.7%) had detailed advanced T-stage (p=0.002), lymph node metastases (p=0.009), advanced follow-up. Pathologic staging was T2N0, T3N0, T2N2, T3N2, T4N2, and intraoperative-pathological stage (p<0.001) and age ≥ 70 years (p=0.036) T4N3 in 6(25%), 4(20%), 2(10%), 8(33%), 1(5%), and 1(5%), respectively. were associated with significant inferior survival in the univariate analyses. 24 patients underwent treatment of 107 lesions with a median disease- Histology, gender, type of additional resections, type of recurrence and free interval prior to cryoablation of 24.5 months. 19/24 patients (79.2%) laterality had no significant impact on survival. Macroscopic complete underwent multiple cryoablations with a mean of 4.25+5.24 (range 1-25) resection remained the only significant prognostic factor in the multivariate lesions/patient. Lesions measured a mean of 32.5+15.9 mm (range 9-113) analysis. by 18.1+8.5 mm (range 6-60) in diameter. Tumor volumes were calculated (0.4XLXW2) at a mean of 7205+16798 mm3 (range 172.8-136,730). Lesion Conclusion: Lung-sparing RP within a trimodality therapy concept is were treated with a mean of 1.74+0.74 probes (range 1-4) and to a mean associated with promising long-term survival, morbidity and mortality. of 2.73+0.85 freeze-thaw cycles (range 2-4). The mean maximum freezing Patients aged ≥ 70 years should be selected very carefully for trimodality time was 9.69+1.09 minutes (range 7-10) with a mean total freeze time therapy. MCR is the most important prognostic factor within this of 24.1+7.98 minutes/lesion (range 18-40). The mean maximum thaw trimodality approach. High rate of locoregional failure warrants further time between freeze cycles was 6.65+1.88 minutes (range 2-10). Extent investigation of locoregional control of the disease. of freezing was monitored by CT imaging with “ice balls” measuring a mean 6.64+2.26 mm (range 0-12.6) beyond the radiographic edge of the Disclosure: No significant relationships. tumor. The procedural morbidity was low and consisted of hematoma, small pneumothorax, hemoptysis and chest pain in one patient each and erythema in 2 chest wall subcutaneous lesions (6/107 =5.6%). All patients were treated as outpatients. 102/105 cryoablated lesions (95.3%) were completely controlled following a single cryoablation treatment as measured by serial PET scans. Failures were due to 1) incomplete treatment due to inexperience in the initial patient, 2) late central recurrence in large area in two patients (3 failures), and 3) recurrence due to anatomic location challenges in 1 patient. Following the initial ablation, median survival was 11.4 months. Overall median survival with this strategy was 36.1 months, and 10/24(41.7%) patients were still alive at a median of 48.8 mos following surgery.

iMig2012.org • Abstract Book 24 Conclusion: Cryoablation for localized recurrent malignant pleural Gemcitabine Cisplatin N # Pts with Grade 3-5 SAEs mesothelioma following surgery can be performed safely as an outpatient Dose Dose procedure with minimal morbidity (5.6%), a very high efficacy (95.3%), and impressive overall survival (36.1 mos). 100 0 9 0 100 225 11 1 (Renal) Disclosure: No significant relationships. 200 225 9 3 (Thrombosis, Abdominal pain, Renal) 300 225 5 1 (Renal)

SESSION IIB mULTI-MODALITY 400-700 175 24 0 September 12, 2012 14:20-16:00 800 175 6 1 (Pulmonary embolism) IIB.4: PROSPECTIVE PHASE I TRIAL OF EXTRAPLEURAL 900 175 6 0 PNEUMONECTOMY OR PLEURECTOMY/DECORTICATION, 1000 175 18 3 (Leukopenia (2), DVT) september 12, 2012 INTRATHORACIC/INTRAPERITONEAL HYPERTHERMIC [IOHC] 1100 175 6 3 (Leukopenia (2), Renal) CISPLATIN AND GEMCITABINE WITH INTRAVENOUS AMIFOSTINE AND SODIUM THIOSULFATE CYTOPROTECTION FOR PATIENTS Conclusion: This prospective study demonstrates the feasibility and WITH RESECTABLE MALIGNANT PLEURAL MESOTHELIOMA safety of administering hyperthermic intraoperative intracavitary combination chemotherapy with cisplatin and gemcitabine following David J. Sugarbaker1, Marcelo Dasilva2, Jeffry Supko3, Olivia Winfrey1, cytoreductive surgery for MPM. It establishes MTD for these drugs, Hannah Eisen1, Juliann Barlow1, Raphael Bueno1, William G. Richards2 respectively, at 175 and 1000 mg/m2. Morbidity and mortality were 1Division Of Thoracic Surgery, Brigham And Women’s Hospital, Boston/ acceptable. A pharmacokinetic gradient was demonstrated that permits MA/UNITED STATES OF AMERICA, 2Division Of Thoracic Surgery, Brigham high concentrations of intracavitary gemcitabine while minimizing systemic And Women’s Hospital And Harvard Medical School, Boston/MA/UNITED toxicity. Early survival analysis is encouraging. STATES OF AMERICA, 3Clinical Pharmacology, Massachusetts General Hospital, Boston/MA/UNITED STATES OF AMERICA Disclosure: No significant relationships.

Background: We sought to determine the maximum tolerated dose (MTD) of gemcitabine added to cisplatin intra-operative heated chemotherapy (IOHC) following either extrapleural pneumonectomy or pleurectomy/ SESSION IIB mULTI-MODALITY decortication as a treatment for mesothelioma. We also investigated September 12, 2012 14:20-16:00 the toxicity profile, perioperative mortality and pharmacokinetics of this treatment. IIB.5: RESULTS OF SHORT ACCELERATED HYPOFRACTIONATED HEMITHORACIC INTENSITY MODULATED RADIATION THERAPY Methods: Between November 2007 and October 2011, 104 patients FOLLOWED BY EXTRAPLEURAL PNEUMONECTOMY FOR underwent surgical resection followed by HIOC with cisplatin (225 mg/ MALIGNANT PLEURAL MESOTHELIOMA m2; previously determined MTD), and dose- escalated gemcitabine. For each surgery type, three patients were enrolled per dose level. Escalation Marc De Perrot, Isabelle Opitz, Masaki Anraku, Victoria Ford, Natasha occurred if there were no dose-limiting toxicities (DLT), defined as grade Leighl, Ronald Feld, John Cho 3 or higher toxicity relatable to study treatment. The concentration of Toronto General Hospital And Princess Margaret Hospital, Toronto/ON/ gemcitabine in perfusate and plasma samples was determined by high CANADA performance liquid chromatography with tandem mass spectrometric detection. Background: The surgical treatment of malignant pleural mesothelioma international mesothelioma interest group (MPM) remains controversial. We and others have observed that MPM Results: Median age was 65 (43-85) and 22 (21%) were women. Histology are radiosensitive and that hemithoracic intensity modulated radiation was epithelial (63), Biphasic (32), and Sarcomatoid (8). Two patients therapy (IMRT) is well tolerated in the adjuvant setting after extrapleural died perioperatively (2%). Toxicity data are given by gemcitabine dose pneumonectomy (EPP). We, therefore, developed a new protocol with (Table). Due to renal toxicity observed with low-dose gemcitabine, neoadjuvant hemithoracic IMRT followed by EPP. The potential advantages 2 the cisplatin dose was reduced to 175 mg/m . The DLT was Grade 3 of this protocol were optimal delivery of the radiation in the preoperative leukopenia, observed in two patients at a gemcitabine dose of 1100 mg/ setting and reduced risk of viable tumor spillage during the surgery due to 2 2 m , establishing the gemcitabine MTD at 1000 mg/m , with 175 mg/ the tumoricidal action of radiation. m2 of cisplatin. The average ± SD maximum concentration of gemcitabine measured in the perfusate was 327 ± 126 µg/mL in patients receiving a Methods: Patients were eligible if they had clinically resectable T1-3N0M0 2 500 mg/m dose and 688 ± 221 µg/mL in patients treated with 1,000 mg/ MPM. Patients received 25 Gy in 5 daily fractions over 1 week to the 2 m . Systemic absorption of gemcitabine was very low, as indicated by the entire ipsilateral hemithorax by IMRT with concomitant boost of 5 Gy to average peak concentrations of gemcitabine in plasma, which were only gross disease based on CT and PET scan findings. EPP was performed one 0.23 ± 0.13% of the maximum drug concentration in perfusate in patients week after the end of radiation. Adjuvant chemotherapy was offered to 2 evaluated at the 500 mg/m dose level and 0.35 ± 0.26% at the 1,000 mg/ patients with N2 disease on final pathology. The primary end-point was 2 m dose level. Fifty-four patients remain alive with a median follow-up the proportion of patients treated without treatment related mortality. An interval of 12.5 months. Estimated median overall survival is 26.6 months accrual of 12 patients was planned for the feasibility study with extension for epithelial, 12.2 months for biphasic, and 8.8 months for sarcomatoid to a phase II study if there were less than 3 treatment related deaths histology. within 30 days of surgery. Toxicities were graded according the CTCAE v4.0 criteria. Patients were followed clinically and radiologically with CT scans.

Results: A total of 20 patients (17 males, 64±7 years old, 17 right sided tumors) were included in this study between 11/2008 and 04/2012. All patients completed IMRT and EPP. IMRT was well tolerated with no grade 3-5 toxicity. EPP was performed 6±3 days after completion of IMRT. No patients died within 30 days of surgery or in-hospital. During follow- up, one patient died from empyema at 88 days (Table 1). Pathological stage was ypT3-4N0M0 (n=8), ypT3-4N2M0 (n=10), ypT4N3M0 (n=1) and ypT3N2M1 (n=1). Four patients received adjuvant chemotherapy for ypN2-3. After a median follow-up of 11 (range 3-42) months, 7 patients developed

iMig2012.org • Abstract Book 25 recurrence, all with ypN2-3 disease. Recurrence were located in the Results: A total of 121 patients were identified on an “intent to treat” basis abdomen (n=3), ipsilateral chest wall (n=3), contralateral lung (n=2) and/or between 1997 and 2011. 94 (77.7%) were male while 27 (22.3%) were pericardium (n=1). The disease-free survival (DFS) reached 52% at 2 years female. 80 (66.2%) had right-sided tumors while 41 (33.9%) were left. and was significantly better in patients with ypN0 disease (100% DFS at 2 Mean age was 65.9 years (range 27-84) with a median age of 68 (male) years vs 20% in the remaining patients; p=0.007). Table 1. Complications and 56 years (female). 40 (33.1%) were >70 years old. Preoperative clinical occurring after induction hemithoracic IMRT and EPP staging was stage I, II, III, and IV in 109 (90.1%), 1, (0.8%), 7 (5.8%), and 4 (3.3%), respectively. Pathologic T stage was T2 in 24 (19.8%), T3 in 70 (57.9%), and T4 in 27 (22.3%); while N stage was N0 in 57 (47.1%), N1 in 3 (2.5%), N2 in 58 (47.9%), and NX in 3 (2.5%). Yielding a pathologic Grade Grade Grade Grade Grade Grade stage I, II, III, and IV in 16 (13.2%), 3 (2.5%), 74 (61.2%), and 28 (23.1%), 0 1 2 3 4 5 respectively. Overall median survival for all 121 patients was 13.8 mos, Thromboembolic 17 0 1 1 1 0 while significantly better survival was noted among female patients event (20.7 vs 12.1 mos), nonsmokers (16.3 vs 11.9 mos), patients without an Atrial fibrillation 14 0 2 4 0 0 identifiable asbestos exposure history (23.4 vs 13.4 mos), and patients without lymph node involvement (N0=20.2 mos vs N1=14.9 mos vs september 12, 2012 Wound infection 17 0 2 1 0 0 N2=9.8 mos vs NX=4.2 mos) (p<0.05). Median survival for epithelioid Chylothorax 18 0 0 2 0 0 histology (17.8 mos) was significantly better than both biphasic (10.3 mos) and sarcomatoid (2.1 mos) subtypes (p<0.01). The median survival Hemothorax 19 0 0 0 1 0 of patients completing standard surgical and adjuvant therapy, i.e., Wound dehiscence 16 1 2 1 0 0 pleurectomy/decortication and radiation with delayed or no postoperative chemotherapy (85 patients=70.2%), was 19.7 mos which equals that Diaphragm patch 19 0 0 0 1 0 reported for trimodality therapy for similar patient groups (median survival dehiscence = 19.0 mos; Sugarbaker, et al; J Thorac Cardiovasc Surg 1999;117(1):54-65), Renal dysfunction 19 0 0 1 0 0 particularly when compared to neoadjuvant chemotherapy followed by EPP Empyema 19 0 0 0 0 1 + hemithoracic radiation (median survival = 16.8 mos; Krug, et al; J Clin Oncol 2009;27(18):3007-13 and median survival = 14.4 mos; Treasure, et Bronchopleural 20 0 0 0 0 0 al Lancet Oncol 2011;12(8):763-72). fistula Conclusion: The results using chemotherapy given in a delayed fashion Conclusion: Short accelerated hypofractionated hemithoracic IMRT at the time of 1st recurrence following lung-sparing pleurectomy/ followed by EPP is feasible and safe. This combination therapy could decortication revealed favorable patient outcomes comparable or better provide improved outcome in resectable MPM patients with ypN0 disease. to those reported for “trimodality” therapy including the recent MARS trial. This suggests that a more rational and conservative approach to Disclosure: No significant relationships. multimodality treatment of patients with malignant pleural mesothelioma may be warranted.

Disclosure: No significant relationships. SESSION IIB mULTI-MODALITY September 12, 2012 14:20-16:00

IIB.6: THE TIMING OF CHEMOTHERAPY IN THE MULTIMODALITY SESSION IIB mULTI-MODALITY TREATMENT OF MALIGNANT PLEURAL MESOTHELIOMA September 12, 2012 14:20-16:00

Robert B. Cameron1, Olga M. Olevsky2, Michael Selch3, Michael IIB.7: PHOTODYNAMIC THERAPY (PDT) AS AN INTRAOPERATIVE international mesothelioma interest group Fishbein4, Fereidoun Abtin5, W Dean Wallace4, Chi Lai4, Robert Suh5, ADJUVANT FOR MALIGNANT PLEURAL MESOTHELIOMA Anne Rorie1 1Thoracic Surgery, David Geffen School Of Medicine At UCLA, Los Angeles/ Joseph Friedberg CA/UNITED STATES OF AMERICA, 2Division Of Hematology-Oncology, David University Of Pennsylvania/UNITED STATES OF AMERICA Geffen School Of Medicine At UCLA, Santa Monica/CA/UNITED STATES OF AMERICA, 3Radiation Oncology, David Geffen School Of Medicine At UCLA, Photodynamic Therapy (PDT) is a light-based cancer treatment that Los Angeles/UNITED STATES OF AMERICA, 4Pathology And Laboratory is particularly well suited as an intraoperative adjuvant treatment for Medicine, David Geffen School Of Medicine At UCLA, Los Angeles/CA/ malignant pleural mesothelioma. With patient education and standardized UNITED STATES OF AMERICA, 5Radiology, David Geffen School Of Medicine dosimetric techniques PDT can be combined safely and easily with surgery. At UCLA, Los Angeles/CA/UNITED STATES OF AMERICA Our group has published some encouraging results on the combination of PDT and radical pleurectomy where the 31 epithelial subtype patients Background: Various strategies have been used with regard to the timing (100% IMIG Stage III/IV) had a median survival from the time of surgery of chemotherapy within a multimodality treatment approach to malignant of 41.2 months. Our hypothesis is that PDT played a role in these unusual pleural mesothelioma. Previously, postoperative chemotherapy has been results. Beyond the fact that PDT treats tissue for several millimeters below evaluated but recently, neoadjuvant chemotherapy has been more popular. the surface, It has the additional advantages of being compatible with We evaluated a strategy of delayed chemotherapy given only at the time of essentially any other treatment modality and has potential to be synergistic a documented recurrence following our standard protocol of pleurectomy/ with a number of them. Future directions for our group include performing decortication followed by adjuvant radiation. a randomized Phase III study to definitively test the hypothesis that PDT is contributing to survival. In addition we are also performing translational Methods: Following IRB approval, we retrospectively reviewed our research aimed at improving our current approach to intraoperative PDT prospective thoracic database to identify patients treated with a while at the same time developing new treatments designed to capitalize basic protocol including lung-sparing pleurectomy/decortication upon the known immunostimulatory effects of PDT. followed by adjuvant radiation therapy. Patients were then followed for disease recurrence and only treated with chemotherapy at the time of Disclosure: No significant relationships. identification of at least one indicator lesion (minimal recurrent disease). Patient characteristics, results, and survival were assessed and compared to those reported in the literature for trimodality therapy with extrapleural pneumonectomy (EPP).

iMig2012.org • Abstract Book 26 Session IIC Immunology September 12, 2012 14:20-16:00

SESSION IIC IMMUNOLOGY SESSION IIC IMMUNOLOGY September 12, 2012 14:20-16:00 September 12, 2012 14:20-16:00 september 12, 2012 IIC.1: MEASLES VIRUS VACCINE INDUCES ONCOLYSIS OF IIC.2: DENDRITIC CELL BASED IMMUNOTHERAPY IN MESOTHELIOMA CELLS AND ALLOWS DENDRITIC CELLS TO COMBINATION WITH METRONOMIC CYCLOPHOSPHAMIDE IN ACTIVATE TUMOR-SPECIFIC CD8 RESPONSES PATIENTS WITH MESOTHELIOMA

Marc Gregoire1, Jean-François Fonteneau1, Nicolas Boisgerault1, Jean- Joachim G. Aerts1, Robin Cornelissen2, M E.H. Lambers3, H C Baptiste Guillerme1, Frédéric Tangy2 Hoogsteden2, Joost P.J.J. Hegmans1 1INSERM, Nantes/FRANCE, 2Pasteur Institute, Viral Genomics and 1Pulmonary Diseases, Erasmusmc, Rotterdam/NETHERLANDS, 2Pulmonary Vaccination, Paris/FRANCE Medicine, Erasmus Mc, Rotterdam/NETHERLANDS, 3Pulmonary Medicine, Erasmus Medical Centre, Rotterdam/NETHERLANDS Despite conventional medical and surgical treatments, malignant pleural mesothelioma (MPM) remains incurable. Thus, other therapeutic Background: Dendritic Cells (DC) are extremely potent antigen presenting options must be evaluated. Oncovirotherapy (i.e., the use of replication- cells capable of inducing a CD8+ cytotoxic T cell reaction. In an earlier competent virus for cancer treatment) is currently explored in clinical study on autologuous tumor lysate pulsed DC based immune therapy trials. We recently investigated the antineoplastic potential of a live- in mesothelioma (MM) we showed that we were able to induce an attenuated measles virus (MV) strain, usually used as vaccine since more anti-tumor response. However it is known that the induced anti-tumor than 60 years. We evaluated both oncolytic activity and immunoadjuvant immunity in cancer patients is created in a unfavorable highly suppressive properties of the MV vaccine strain on mesothelioma cells. Infectivity, tumor environment. Regulatory T-cells (Tregs) are prominent cells in this syncytium formation and cytolytic activity of MV were studied on a suppressive environment. We have previously showed the presence of panel of mesothelioma cell lines derived from pleural effusions of MPM Tregs in patients with MM. In a murine model on MM, depletion of Treg patients. We observed that MV infected preferentially MPM cell lines in with metronomic cyclophosphamide (CTX) could enhance the anti-tumoral comparison with non-transformed cells, leading to an efficient killing immune response induced by DC vaccination1. of a significant fraction of tumor cells. A cytoreductive activity was also evidenced through formation of multinuclear cellular aggregates (syncytia). Methods: A non-randomized study was performed in patients with MM We also evidenced that MV infection of mesothelioma cancer cells elicited who were non progressive after standard chemotherapy. A pleurectomy/ an autologous antitumor immune response. We showed that MV vaccine decortication (P/D) was performed before DC vaccination in case that was induced apoptotic cell death of infected tumor cells. The apoptotic considered best interest of the patient. According to our previous study, 3 infected cells were thus phagocytosed by monocyte-derived (Mo-DC) doses of pulsed autologuous DC were re-injected every 2 weeks. Patients [Cancer Res 2008;68(12):4882–92] and plasmacytoid (pDC) dendritic cells were treated with 100mg CTX/day for 7 days starting 9 days before every

[submitted]. Loading of these Mo-DC and pDCs with MV-infected MPM vaccination. After the 3rd vaccination again 7 days of CTX were taken. international mesothelioma interest group cells induced spontaneous DCs maturation, as evidenced by the increased expression of MHC and costimulatory molecules along with the production Results: 9 patients were enrolled. Mean age 60 (range 35-78yrs). Data of proinflammatory cytokines. Priming of autologous T cells by the two on Treg levels will be presented at the conference. Respone according to types of DCs loaded with MV-infected MPM cells led to a significant modified RECIST and survival are presented in the table proliferation of tumor-specific CD8 T cells. Since the pleural cavity is a confined compartment that could allow an efficient interaction between cancer cells and a viral therapeutic agent, this constitutes an ideal target for the local administration of high virus concentrations. This intrapleural administration pathway could also be a solution to limit virus inactivation by circulating MV neutralizing antibodies. Altogether, these data strongly support the potential of oncolytic MV as an efficient therapeutic agent for mesothelioma.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 27 was analyzed by intracellular cytokine staining using flow cytometry. To

block iNKT cell activation, mice were treated with 0.5mg of anti-CD1d- No Response to Pleurectomy/ Response to DC Survival blocking antibody 1 day before and 7 days after tumor cell injection. To chemotherapy Decortication immunotherapy (Months) activate iNKT cells, αGarCel was injected intraperitoneally on day 1, 5 and 9 after the tumor cell injection and the survival, the amount of pleural 1 SD No SD 24 effusion on day 14 were analyzed. IFN-gamma expression on iNKT cells and CD8 positive T cells were also analyzed by flow cytometry.

2 SD No SD 25 Results: The ratio of iNKT cells to CD3 positive cells in the pleural cavity significantly increased after tumor cell injection (day 0: 0.8±0.2%, day 7: 7.8±0.6%, day 14: 11.7±0.6%) while that in spleen did not change. 3 SD No SD 14 The ratio of iNKT cells to CD3 positive cells in the tumor on day14 was 8.9±1.0%. These iNKT cells in the pleural cavity and the tumor showed higher CD25 expression compared with that in the spleen. The ratio of IFN- 4 PR No CR 12 alive gamma positive iNKT cells in the pleural cavity was significantly increased september 12, 2012 after the tumor cell injection (day 0: 3.0±0.9%, day 7: 11.4±1.3%, day 14: 46.3±5.4%). The mice treated with CD1d-antibody showed less CD25 5 SD No SD 14 alive expression on iNKT cells, less IFN-gamma positive iNKT cells in the pleural cavity and increased amount of pleural effusion on day 14 compared

with the control mice (1.1±0.1ml and 0.7±0.1ml, respectively, p=0.045). 6 SD Yes SD 20 The mice treated with αGalCer showed significantly prolonged survival compared with the control mice (median survival time, 23.0 days and 17.0 7 PR Yes PD 12 days, respectively, p < 0.0001) and was associated with increased ratio of IFN-gamma positive iNKT cells and CD8 positive T cells in the pleural effusion. 8 SD Yes PR 26 alive Conclusion: iNKT cells appear to contribute to the anti-tumor immune response in murine MPM. Modulation of iNKT cells could be a new 9 PR Yes SD 37 alive therapeutic approach for patients with MPM.

Disclosure: No significant relationships. SD = stable disease, PR = partial response, CR = complete response, PD = progressive disease. No major side effects were determined during the study SESSION IIC IMMUNOLOGY Conclusion: CTX can be safely administered during DC immunotherapy. September 12, 2012 14:20-16:00 The results on Tregs will be presented at the conference. The results on overall survival confirm the data found from our earlier study on DC IIC.4: OPTIMISING REGULATORY T CELL (TREG) DEPLETION IN immunotherapy in patients with mesothelioma. Hegmans, Veltman, Lambers, de Vries, Figdor, Hendriks, Hoogsteden, Lambrecht, Aerts. Am J COMBINATION WITH CHEMOTHERAPY FOR ENHANCED ANTI- Respir Crit Care Med 2010;181:1282. TUMOUR IMMUNITY

1 2 2 Disclosure: No significant relationships. Alistair M. Cook , Alison M. Mcdonnell , Bruce W.S. Robinson , Michael J. Millward1, Richard Lake2, Anna K. Nowak2 1School Of Medicine And Pharmacology, University Of Western Australia, international mesothelioma interest group Perth/WA/AUSTRALIA, 2School Of Medicine And Pharmacology, National SESSION IIC IMMUNOLOGY Centre For Asbestos Related Diseases, University Of Western Australia, September 12, 2012 14:20-16:00 Perth/WA/AUSTRALIA

IIC.3: ANTI-TUMOR ROLE OF INTERFERON-GAMMA PRODUCING Background: Cytotoxic chemotherapy is widely used to palliate malignant CD1D-RESTRICTED NKT CELLS IN MURINE MALIGNANT PLEURAL pleural mesothelioma (MM) and non small cell lung cancer (NSCLC). While originally considered detrimental to the immune system, there is MESOTHELIOMA now abundant preclinical data showing that chemotherapy can enhance anti-cancer immunotherapy. Tregs are immunosuppressive CD4+ T cells Tetsuzo Tagawa, Licun Wu, Zhihong Yun, Katrina Rey- Mcintyre, thought to inhibit anti-tumour immune responses; murine data suggests Marc De Perrot that Treg eradication may augment existing anti-tumour immunity. Latner Thoracic Surgery Research Laboratories, Toronto General Research Cyclophosphamide (CTX) is immunostimulatory and at low doses Institute, University Of Toronto, Toronto/ON/CANADA selectively depletes Tregs in mice and humans. The primary objective of this study is to identify an optimum dose and schedule of iterative low Background: CD1d-restricted invariant NKT (iNKT) cells can provide dose oral CTX for Treg depletion in the context of pemetrexed-based adjuvant activity against cancer by producing large amounts of IFN-gamma chemotherapy, and to determine how treatment affects the function and which activate other immune cells, and orchestrate protective anti-tumor phenotype of the cellular immune response. immunity. Recently, induction of the iNKT cell-dependent anti-tumor immune response using its ligand alpha-galactosylceramide (αGalCer) has Methods: This single centre phase 1b study enrolled patients with been attempted in several tumor types. However, the role of iNKT cells in advanced MM or NSCLC planned for pemetrexed based chemotherapy. The the tumor microenvironment has not yet been fully addressed. Our aim is first 11 patients received pemetrexed 500 mg/m2 ± cisplatin 75 mg/m2 or to elucidate the role of iNKT cells in the thoracic cavity by using a murine carboplatin AUC 4-6 on a standard 21 day schedule, and from the second malignant pleural mesothelioma (MPM) model. cycle received escalating doses of oral CTX taken every day for days 1-14 of each cycle, with an initial dose of 50 mg daily (‘intermittent’ group). All Methods: Half a million of AB12 murine malignant mesothelioma cells subsequent patients received oral CTX continuously from the second cycle were injected into the right pleural cavity of female Balb/c mice. Mice (‘continuous’ group). CTX dose was increased in subsequent cycles until were sacrificed and the pleural effusion and tumors were collected on Tregs comprised <4% of the CD4+ T cell population. Weekly peripheral days 0 (before tumor cell injection), 7 and 14 after tumor cell injection. blood samples were collected, and the CD25+CD127loFoxp3+ proportion of Lymphocytes were isolated and iNKT cells were identified using CD3 CD4+ T cells (‘Treg%’) in whole blood determined by flow cytometry in real antibody and CD1d tetramers loaded with αGalCer. Cytokine expression

iMig2012.org • Abstract Book 28 time. Peripheral blood mononuclear cells (PBMC) were cryogenically stored SESSION IIC IMMUNOLOGY for subsequent analysis. Toxicity and radiological response were assessed. September 12, 2012 14:20-16:00

Results: To date, 26 patients with MM (21/26) or NSCLC (5/26) have IIC.6: ANTI-TUMOR ACTIVITY INDUCED BY CTLA-4 completed treatment. Preliminary analyses demonstrate the Treg% is BLOCKADE MIGHT BE MEDIATED BY NKT CELLS IN A MURINE stable during the first cycle of standard care chemotherapy, with a mean MESOTHELIOMA MODEL pre-treatment baseline of 8.08% ± 2.08%. Following commencement of CTX, the peripheral blood Treg% decreased (with optimal Treg Licun Wu, Zhihong Yun, Yidan Zhao, Marc De Perrot depletion achieved during treatment with alternating 50/100 mg per Latner Thoracic Surgery Research Laboratories, Toronto General Research day) then increased back to/and above baseline during the treatment Institute, University Of Toronto, Toronto/ON/CANADA break. In ‘intermittant’ patients, the Treg nadir was 5.90% ± 1.51%, and was achieved on cycle 3 day 15. ‘Continuous’ patients did not receive a Background: Considerable evidence has shown that cancer treatment break, in an effort to negate the Treg% increase; however this immunotherapy is promising when combined with chemotherapy. did not produce a difference in Treg% when compared to the ‘intermittant’ Immunosuppresive components such as regulatory T cells (Treg), cytotoxic group. Increasing doses up to alternating 100/150 mg daily did not result in T cell associated antigen-4 (CTLA-4), and so on, are major hurdles affecting september 12, 2012 improved Treg depletion. Analysis of PBMC T cell populations indicates that the immune surveillance in tumor microenvironment. Therefore, removal of cell proliferation (Ki67) and activation (inducible co-stimulator; ICOS) peak these brakes would be able to enhance the anti-tumor immune reaction, and trough with each cycle of chemotherapy, declining during treatment thus improve the efficacy of chemotherapy in mesothelioma. Our previous and increasing during the treatment break. Combining CTX with routine studies indicated that Treg depletion or blockade of CTLA-4 signalling cytotoxic chemotherapy is feasible with no additional haematological or between cycles of chemotherapy improved the outcome of mesothelioma. other toxicities. We notice that the number of NKT cells increased over time at the early stage of tumor development. Tumors grew more rapidly in CD1d KO mice Conclusion: The addition of CTX to pemetrexed based chemotherapy is than WT mice, therefore NKT cells may play important roles in mediating safe and feasible. Doses above alternate days 50/100 mg do not improve anti-tumor effect. NKT cell activation might be a potent approach to depletion. Additional work will investigate the effect of this chemo- mesothelioma treatment. immunotherapy protocol on tumour-specific cellular immunity. Methods: The effect on tumor growth was evaluated in subcutaneous Disclosure: No significant relationships. murine mesothelioma model. CTLA-4 blocking antibody +/- NKT ligand α-GalCer was administered following each cycle of chemotherapy in WT or CD1d KO Balb/c mice, and monotherapy was included as controls. Anti-tumor effect was evaluated by tumor growth delay and survival of SESSION IIC IMMUNOLOGY the animals. Tumor cell repopulation was quantified by BrdU incorporation September 12, 2012 14:20-16:00 and Ki67 by immunohistochemistry and/or flow cytometry. NKT cells were identified by α-GalCer conjugated tetramer staining, and RT-PCR was IIC.5: GEMCITABINE AND CTLA-4 BLOCKADE SYNERGIZE IN THE performed to determine the gene expression of associated cytokines. INDUCTION OF ANTI-MESOTHELIOMA IMMUNITY Results: Anti-tumor effect was achieved by administration of CTLA- W J. Lesterhuis, Joanne Salmons, Bruce W.S. Robinson, 4 blockade or α-GalCer between cycles of chemotherapy. Tumor cell Anna K. Nowak, Richard Lake repopulation during the intervals of cisplatin was significantly inhibited. School Of Medicine And Pharmacology, University Of Western Australia, Anti-CTLA-4 therapy gave rise to an increased number of CD4 and CD8 T Perth/WA/AUSTRALIA cells infiltrating the tumor. RT-PCR demonstrated that the gene expression of IL-2, IFN-γ, granzyme B, and perforin increased in the tumor milieu. NKT Background: Several chemotherapeutics exert immunomodulatory effects. cell activation by α-GalCer had mild effect, but α-GalCer combined with One of these is the nucleoside analogue gemcitabine, which is used in cisplatin was more effective inhibiting tumor growth than other groups international mesothelioma interest group patients with mesothelioma, but with modest efficacy. We hypothesized in WT mice, whereas α-GalCer did not result in additional effect in CD1d that the immunopotentiating effects of this drug are partly restrained by KO mice. Interestingly, α-GalCer plus CTLA-4 blockade resulted in more the inhibitory T cell molecule CTLA-4 and thus could be augmented by IFN-γ production in WT mice rather than CD1d KO mice during cisplatin combining it with a blocking antibody against CTLA-4. treatments.

Methods: Balb/c AB1-HA mesothelioma-bearing mice were treated with Conclusion: Blockade of CTLA-4 signalling demonstrated effective anti- gemcitabine or cisplatin, with or without a CTLA-4 blocking antibody in a tumor effect correlating with inhibiting cancer cell repopulation between concurrent or consecutive schedule. Immune responses were monitored in cycles of chemotherapy. This effect might be mediated by NKT cells in WT the tumor, tumor-draining lymph nodes and non-draining lymph nodes. mice. For mechanistic information, in separate studies CD4 and CD8 cells were depleted. Disclosure: No significant relationships.

Results: We found that treatment with gemcitabine chemotherapy in combination with anti-CTLA-4 blockade results in the induction of a potent anti-tumor immune response. Mice treated with the combination exhibited SESSION IIC IMMUNOLOGY rapid and clear mesothelioma regression and long-term protective September 12, 2012 14:20-16:00 immunity. This was not the case for cisplatin. In addition, we show that the efficacy of the combination is critically dependent on the timing of IIC.7: OPTIMAL SOURCE OF WHOLE TUMOR ANTIGENS FOR administration of the two agents and is CD4 and CD8 T cell-dependent. DENDRITIC CELL-BASED IMMUNOTHERAPY IN MURINE MESOTHELIOMA Conclusion: This is the first demonstration of a clear synergistic effect of classic cancer chemotherapy and immune checkpoint blockade. These Joost P.J.J. Hegmans, M E.H. Lambers, N Mahawi, Jacqueline Dekkers, results provide a basis to pursue combination therapies with anti-CTLA-4 H.C. Hoogsteden, Joachim G. Aerts and immunopotentiating chemotherapy in mesothelioma and have Pulmonary Diseases, Erasmusmc, Rotterdam/NETHERLANDS important implications for future studies. Since both drugs are approved for use in patients our data can be immediately translated into clinical Background: Malignant mesothelioma is an aggressive tumor which trials. is resistant to conventional therapies. This study focuses on dendritic cell (DC) vaccination for malignant mesothelioma which is a novel and Disclosure: No significant relationships. promising strategy in cancer treatment. The source of tumor associated

iMig2012.org • Abstract Book 29 antigens (TAA) necessary for DC stimulation might influence the level of the anti-tumor responses induced. The aim of this study was to select the optimal TAA source for DC vaccine development for the treatment of malignant mesothelioma. As a second objective, the effectiveness of tumor exosome and apoptotic tumor cell fragments based DC vaccines were compared with the conventional necrotic tumor lysate loaded DC vaccines in vivo.

Methods: Tumor derived exosomes were obtained by ultracentrifuging supernatant of the mouse AB1 mesothelioma cell line culture. For the apoptotic tumor cell fragments, AB1 tumor cells were exposed to 25kJ/ m2 ultraviolet-B light while tumor cells for necrotic material were subjected to freeze-thaw cycles. Twenty-four BALB/c mice were inoculated intraperitoneally with a lethal dose of AB1 cells that leads to terminal illness between 15 and 30 days. For immunotherapy, dendritic cells were cultured september 12, 2012 ex vivo and stimulated with tumor-derived exosomes, apoptotic tumor cells or necrotic tumor cell lysate, and vaccinated at day 7 after tumor inoculation.

Results: Electron microscopy of tumor exosomes, necrotic and apoptotic tumor cell fractions used for vaccine preparation revealed the morphological differences between the fractions. The effectiveness of uptake of the tumor fractions and subsequent DC stimulation was examined by fluorescence microscopy and flow cytometric analysis of DC maturation markers. Our results showed the highest uptake of the apoptotic material and increased expression levels of cell surface markers MHCII, CD40, CD80 and PDL1 after stimulation. Mice receiving a DC vaccine based on tumor derived exosomes, 7 days after tumor cell injection, had a doubled survival rate (33.3% of mice were alive after 52 days) compared to mice receiving the conventional vaccine (16.6% of mice were alive after 52 days). Mice that received a DC vaccine based on apoptotic tumor cell fragments had a survival rate that was three times higher (50% of mice were alive after 52 days) compared to the conventional necrotic vaccine. Tumor material present in three mice after 52 days, had increased activation of specific anti-tumor immunity compared to mice without DC- treatment.

Conclusion: Stimulation of DCs with tumor exosomes, apoptotic or necrotic tumor lysate showed an increased expression of cell surface markers, indicating maturation and activation of DCs. All different DC vaccinations induced successful anti-tumor responses but tumor derived exosomes and apoptotic tumor cell fragments were most efficient. Histology shows that long term surviving mice have an increased anti- tumor response and activation of the immune system compared to mice international mesothelioma interest group without DC-treatment. These promising results obtained in mice could lead to a refined method for DC-based immunotherapy in patients in the near future.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 30 General Session III Surgical Treatment September 13, 2012 08:00-09:30

GENERAL SESSION III SURGICAL TREATMENT therapy. The role of intraoperative chemotherapeutic agents remains September 13, 2012 08:00-09:30 unproven. september 2012 13, GSIII.3: SURGICAL APPROACHES TO MPM - CON References 1. Balduyck B, Trousse D, Nakas A, Martin-Ucar AE, Edwards J, Waller David A. Waller DA. Ann Thorac Surg. 2010 ;89(3):907-11. 2. Martin-Ucar AE, Nakas A, Glenfield Hospital/UNITED KINGDOM Edwards JG, Waller DA. Eur J Cardiothorac Surg. 2007 ;31(5):765-70 3. Edwards JG, Martin-Ucar AE, Stewart DJ, Waller DA.Eur J Cardiothorac “WHEN NOT TO OPERATE” Surg. 2007;31(5):759-64 4. Flores RM, Pass HI, Seshan VE, Dycoco J, Zakowski M, Carbone M, Bains MS, Rusch VW. J Thorac Cardiovasc Surg. There is no indication to perform any more than palliative surgery in 2008;135(3):620-6 5. Lang-Lazdunski L, Bille A, Lal R, Cane P, McLean non-epithelioid MPM. In the treatment of sarcomatoid MPM , unlike other E, Landau D, Steele J, Spicer J.J Thorac Oncol. 2012 Apr;7(4):737-43 6. cell types,the extent of surgery has no influence on postoperative survival Rusch VW, Giroux D, Kennedy C et al. J Thorac Oncol 2012 (in press) 7. which is around 6 months only [1]. Nakas A, Trousse DS, Martin-Ucar AE, Waller DA.Eur J Cardiothorac Surg. 2008;34(4):886-91. 8.Treasure T, Lang-Lazdunski L, Waller D,Lancet There is no indication to perform Extrapleural Pneumonectomy (EPP) in Oncol. 2011;12(8):763-72. 9. Hillerdal G, Sorensen JB, Sundström S, Riska mediastinoscopy positive MPM. There is no survival benefit in EPP over H, Vikström A, Hjerpe A. J Thorac Oncol. 2008 Nov;3(11):1325-31 10. pleurectomy/decortication in these patients. Median postoperative survival Tilleman TR, Richards WG, Zellos L, Johnson BE, Jaklitsch MT, Mueller J, was around 16 months in case-matched patients [2] Yeap BY, Mujoomdar AA, Ducko CT, Bueno R, Sugarbaker DJ. J Thorac Cardiovasc Surg. 2009 ;138(2):405-11 Median sternotomy should be preferred to lateral thoracotomy for

EPP. This approach results in reduced operating time; less postoperative Disclosure: No significant relationships. analgesia and faster recovery [3].

Extended pleurectomy/decortication (EPD) should be preferred to EPP in most cases. The increased morbidity and mortality after pneumonectomy confer a survival benefit for EPD in most cases [4,5]. The benefit of EPP is limited to the small subgroup of patients presenting in stage I disease [6].

There is no role for open incomplete resection. The combination of the poor prognosis of R2 resection and delayed postoperative recovery after thoracotomy mean this approach cannot be justified [7]. If thoracotomy is performed then macroscopic complete resection must be achieved even if international mesothelioma interest group this requires phrenectomy.

Palliative VATS debulking should be preferred to EPD in stage III MPM. In our experience the additional morbidity and mortality of EPD is not justified in N2 positive disease. VATS with an R2 palliative debulking operation has similar long-term benefits. EPD should be limited to node- negative epithelioid cases.

Systemic chemotherapy should be considered as an alternative to radical surgery in most cases. In the MARS trial control arm a median survival of 19 months was obtained from chemotherapy alone; this exceeded the post-EPP survival [8]. Other studies have reported median survival of 23 months for early stage epithelioid MPM from chemotherapy alone [9] which is comparable to surgey in similar groups [6].

The value of intraoperative chemotherapy remains unproven. Whilst the feasibility of the technique has been established and local control may have been improved there has not been a significant improvement in overall survival [10].

Conclusion: Better understanding of the importance of histological cell – type and disease staging has refined selection for surgery in MPM. Surgical strategy has evolved from the use of extrapleural pneumonectomy as the default operation towards lung sparing surgery. Extended pleurectomy/ decortication with macroscopic complete resection should be favoured in node-negative epithelioid MPM. EPP may be considered in the fit, young patient with stage 1 epithelioid disease. In node-positive or non-epithelioid disease the use of surgery by VATS should be considered in a palliative setting. All surgery for MPM should be in conjunction with systemic

iMig2012.org • Abstract Book 31 Session IIIA Surgery for Mesothelioma September 13, 2012 10:00-11:30

SESSION IIIA SURGERY FOR MESOTHELIOMA without pre-operative chemotherapy at Brigham and Women’s Hospital September 13, 2012 10:00-11:30 between 2001 and 2010. Data for treatment, recurrence and survival

were determined from medical records. A dedicated thoracic radiologist september 2012 13, IIIA.1: EPP OR PD: THE CYTOREDUCTIONIST’S DILEMMA reviewed the post-operative chest CT and/or PET-CT scans to determine sites of recurrence. Time to recurrence was calculated from the date David Rice of resection and estimated by the Kaplan-Meier method. Rates were Thoracic Surgery, MD Anderson Cancer Center, TX/UNITED STATES OF compared using Fisher’s exact test. AMERICA Results: Median age was 62 years. 138 patients (81%) were men. Median Cytoreductive surgery for malignant pleural mesothelioma includes tumor volume was 390 cm3. Histology on final pathology was epithelial for extrapleural pneumonectomy (EPP) and extended pleurectomy 104 patients (61%) and non-epithelial for 67 (39%). No patients received decortication (PD). The goal of either procedure is complete macroscopic pre-operative chemotherapy (CT); 134 (78%) received heated intra- resection of tumor. EPP and PD differ regarding extent of resection, operative chemotherapy (HIOC); 78 (46%) received adjuvant CT and 73 morbidity profile, patterns of recurrence and compatability with adjuvant (43%) received adjuvant radiation therapy (RT). RT was delivered using therapies. By definition, EPP removes the ipsilateral lung, pleurae, a matched electron-photon technique for 32 patients (52%), intensity ipsilateral diaphragm and pericardium. PD removes the above structures modulated RT for 22 (35%) and other 3D conformal techniques for 8 but leaves the lung in situ, and also occasionally the diaphragm and (13%). Median RT dose was 54 Gy. Among the 162 evaluable patients, 120 pericardium. Because depleuralized lung remains local recurrence is higher (74%) developed a recurrence. Median follow-up time was 54 months and following PD. Paradoxically, higher rates of distant recurrence after EPP are median time to recurrence was 12.4 months. Sites of first recurrence are reported. Although EPP is perhaps a more oncologically sound procedure shown in the table. it is associated with higher mortality and negatively impacts quality of life more than PD. Lung preservation may also compromise the ability to deliver hemithoracic radiation, which has been effective in reducing SITE OF N % of % % local recurrence rates to below 15% after EPP. Despite this, retrospective RECURRENCE All Patients of Recurrences of Recurrences comparative studies have failed to demonstrate any survival advantage of from 1997 EPP over PD. It is well known that survival after EPP is highly influenced by report* tumor stage and histology. It is also evident that preoperative staging of Ipsilateral 83 51% 69% 67% MPM is highly inaccurate. A possible strategy to refine patient selection for Hemithorax cytoreductive surgery involves thorough intraoperative staging prior to the (IHT) and/or decision to perform EPP or PD. Patients with non-epithelioid or advanced Mediastinum stage (T4/N2) disease are unlikely to obtain any benefit from EPP and may Abdomen 60 37 % 50 % 50%

be better served with PD, whereas those with N0/1 epithelioid tumors may international mesothelioma interest group potentially benefit from the improved local control offered by EPP. Contralateral 41 25 % 34 % 33% Hemithorax Disclosure: No significant relationships. (CHT) Distant 8 5 % 7 % 8%

SESSION IIIA SURGERY FOR MESOTHELIOMA *Baldini EH, Recht A, Strauss GM, et al. Ann Thorac Surg 1997;63:334- September 13, 2012 10:00-11:30 8 45% of patients experienced recurrences in the IHT, 27% in the mediastinum, 51% in the IHT or mediastinum, and 19% in the IHT or IIIA.2: PATTERNS OF RECURRENCE FOLLOWING EXTRAPLEURAL mediastinum only. The most common sites of recurrences in the IHT were PNEUMONECTOMY (EPP) FOR MALIGNANT PLEURAL chest wall mass (68% of recurrences) and neo-pleural mass (27%); in the mediastinum were lymph nodes (79%) and mediastinal soft tissue (28%); MESOTHELIOMA (MPM) in the abdomen were retroperitoneal adenopathy (40%), ascites (38%), abdominal mass (32%), and peritoneal mass (26%); in the CHT were lung Brian M. Goodman1, Ritu R. Gill2, Olivia Winfrey1, William G. Richards3, nodules (56%) and pleural effusion (49%); and for distant sites were bone Aileen B. Chen4, David E. Kozono4, Raymond H. Mak4, Rapahel Bueno3, (75%) and soft tissue (25%). For patients who developed a recurrence in David J. Sugarbaker1, Elizabeth H. Baldini5 the IHT or mediastinum, the rate was lower for those who received RT 1Division Of Thoracic Surgery, Brigham And Women’s Hospital, Boston/MA/ (29/73, 40%) compared to those who did not receive RT (54/89, 61%; UNITED STATES OF AMERICA, 2Radiology, Brigham And Women’s Hospital, p=0.01). Boston/MA/UNITED STATES OF AMERICA, 3Division Of Thoracic Surgery, Brigham And Women’s Hospital And Harvard Medical School, Boston/MA/ Conclusion: The most common site of recurrence after EPP and planned UNITED STATES OF AMERICA, 4Radiation Oncology, Brigham And Women’s 5 trimodality therapy remains the ipsilateral hemithorax (including Hospital, MA/UNITED STATES OF AMERICA, Radiation Oncology, Brigham mediastinum) and true distant failure remains unusual. The distribution of And Women’s Hospital/Dana-Farber Cancer Institute, Boston/MA/UNITED recurrences is strikingly similar to our prior report from 15 years ago. STATES OF AMERICA Disclosure: No significant relationships. Background: We have previously described patterns of failure following EPP and trimodality therapy for MPM. We sought to update our results with a larger cohort of contemporary patients.

Methods: We reviewed records for 171 patients who underwent EPP

iMig2012.org • Abstract Book 32 SESSION IIIA SURGERY FOR MESOTHELIOMA pleurectomy or extrapleural pneumonectomy, and found superior results September 13, 2012 10:00-11:30 with the radical pleurectomy group. This report summarizes our current results for the expanded cohort of patients having undergone radical IIIA.3: LUNG-SPARING RADICAL PLEURECTOMY IS ASSOCIATED pleurectomy with intraoperative porfimer sodium photodynamic therapy. WITH IMPROVEMENT OF PULMONARY FUNCTION AND LUNG PERFUSION IN PATIENTS WITH MALIGNANT PLEURAL Methods: 44 patients (37-81 years) underwent radical pleurectomy MESOTHELIOMA with intraoperative porfimer sodium photodynamic therapy. The goal of every operation was to achieve a macroscopic complete resection, while Servet Bölükbas1, Michael Eberlein2, Joachim Schirren1 preserving the lung and, whenever possible, the phrenic nerve and as 1Thoracic Surgery, Dr. Horst Schmidt Klinik, Wiesbaden/GERMANY, 2Division much of the pericardium and diaphragmatic musculature as possible. The Of Pulmonary, Critical Care And Occupational Medicine, Carver College Of decision to perform radical pleurectomy was a preoperative decision, not Medicine, University Of Iowa, Iowa/UNITED STATES OF AMERICA intraoperative, regardless of tumor bulk or degree of pulmonary fissure invasion. 38/44 patients also received pemetrexed-based chemotherapy. Background: Pulmonary function is reduced in patients with malignant All survivals were calculated from the time of surgery, not diagnosis or pleural mesothelioma (MPM) due to encassed lung tissue via a rind of other treatments. september 2012 13, tumor with or without concurrent effusion. Re-expansion of the trapped lung might be achieved by radical pleurectomy (RP). The objective of this Results: A macroscopic complete resection was achieved in 43/44 patients study was to investigate changes in pulmonary function and lung perfusion (with the 1 incompletely resected patient undergoing subtype revision in patients undergoing RP. from preoperative epithelial to mixed desmoplastic on the final pathology). There was 1 postoperative mortality (stroke). Average length of stay was Methods: All patients with histologically proven MPM were evaluated for 13.5 days. The median follow-up for all patients was 36.5 months. The trimodality therapy including RP as surgical procedure in a prospective, stage breakdown for the 36 epithelial patients was 1(I)/ 27(III)/8(IV), of nonrandomized study from January to December 2010. Pulmonary- which 24/36 had N2 disease and the median overall survival was 36.6 function tests and perfusion scans were obtained before and 2 months months – 31.7 months for N2 disease and 57.1 months N0-1 disease after RP. Primary end points were pulmonary function (forced vital capacity (p=0.04) . The stage breakdown for the 8 nonepithelial patients was [FVC], forced expiratory volume in 1 second [FEV1]) and ipsilateral lung 6(III)/2(IV), of which 5/8 had N2 disease and the median overall survival perfusion. was 6.8 months.

Results: Sixteen out of 25 consecutive patients were included in the Conclusion: Bearing in mind the limitations of this retrospective series study. Macroscopic complete resection rate was 81.3% (13 patients). there are several sound conclusions that can be drawn from the data. Diaphragm resection was performed in 5 patients (31.3%). Postsurgical This series demonstrates radical pleurectomy can be used to achieve a improvement of PFTs was observed for FVC and FEV1 (both absolute and macroscopic complete resection and that it can be done safely, even in percentage of predicted values) and ipsilateral perfusion (p < 0.001). this cohort of very advanced stage patients (98% stage III/IV) where tumor Avoidance of diaphragm resection was associated with greater increase in volume was often greater than 800 cc. It is also clear that nonepithelial FVC (+34.6±17.0% versus +13.5±5.4%; p = 0.002) and FEV1 (+29.2±18.1% patients do not benefit from this particular approach and it is no longer versus +12.1±6.4%; p = 0.015), respectively. In a linear regression analysis being offered by our group for these patients. The 36.6 month median a lower preoperative FVC (% predicted) or FEV1 (% predicted) was survival from the time of surgery for these advanced stage epithelial associated with higher relative increases in FVC or FEV1 after RP (p < 0.02 patients compares favorably with other surgery-based treatments, for both). especially when matched for stage. Although the epithelial N0-1 patients demonstrated a greater median overall survival of 57.1 months, we do not Conclusion: Lung-sparing RP is associated with significant improvement feel that the 31.7 month median survival for the N2 patients should serve of PFTs and lung perfusion in patients with MPM. Preservation of the as an exclusion criteria for future patients. Overall, we feel these results are diaphragm results in better functional results. Protection of physiological sufficiently encouraging to warrant further study, including a randomized reserve might leave options open for further therapeuty in the long term. phase III to determine if photodynamic therapy is responsible for these international mesothelioma interest group results. Disclosure: No significant relationships. Disclosure: No significant relationships.

SESSION IIIA SURGERY FOR MESOTHELIOMA September 13, 2012 10:00-11:30 SESSION IIIA SURGERY FOR MESOTHELIOMA September 13, 2012 10:00-11:30 IIIA.4: RADICAL PLEURECTOMY AND INTRAOPERATIVE PORFIMER SODIUM PHOTODYNAMIC THERAPY FOR MALIGNANT IIIA.5: INTRAPLEURAL POLYMERIC FILMS LOADED PLEURAL MESOTHELIOMA WITH CISPLATIN FOR MALIGNANT PLEURAL MESOTHELIOMA: PRELIMINARY PHARMACOKINETIC DATA IN AN OVINE MODEL Joseph Friedberg1, Melissa J. Culligan1, Rosemarie Mick2, Stephen M. Hahn3, James Stevenson4, Evan Alley5, Daniel Sterman6, Keith Cengel3 Luca Ampollini1, Stefano Barbieri2, Fabio Leonardi3, Luigi Rolli1, Stefano 1Thoracic Surgery, University Of Pennsylvania, Philadelphia/PA/UNITED Zanichelli3, Antonella Fusari4, Anna Maria Cantoni5, Claudio Mucchino6, STATES OF AMERICA, 2Biostatistics And Epidemiology, University Of Paolo Colombo2, Michele Rusca1, Paolo Carbognani1 Pennsylvania/UNITED STATES OF AMERICA, 3Radiation Oncology, 1Thoracic Surgery, University Hospital Of Parma, Parma/ University Of Pennsylvania/UNITED STATES OF AMERICA,4Taussig Cancer ITALY, 2Dipartimento Farmaceutico, Università Degli Studi Di Parma/ Institute, Cleveland Clinic Foundation, Cleveland/OH/UNITED STATES OF ITALY, 3Clinica Chirurgica Veterinaria E Medicina D’Urgenza, Facoltà Di AMERICA, 5Medicine, University Of Pennsylvania, Philadelphia/UNITED Medicina Veterinaria, Università Degli Studi Di Parma/ITALY, 4Dipartimento STATES OF AMERICA, 6Division Of Pulmonary, Allergy And Immunology, Di Salute Animale, Facoltà Di Medicina Veterinaria, Università Degli Studi University Of Pennsylvania School Of Medicine, Philadelphia/UNITED Di Parma/ITALY, 5Patologia Generale E Anatomia Patologica, Facoltà Di STATES OF AMERICA Medicina Veterinaria, Università Degli Studi Di Parma/ITALY, 6Dipartimento Di Chimica Generale E Inorganica, Chimica Analitica, Chimica Fisica, Background: Our group has conducted several trials for malignant pleural Università Degli Studi Di Parma/ITALY mesothelioma utilizing photodynamic therapy as an intraoperative adjuvant therapy, with porfirmer sodium being the only photosensitizer employed Background: Long-term survival in malignant pleural mesothelioma (MPM) for all patients at a Phase II level. We performed a pilot study comparing patients has been reported after multimodality therapy. Nevertheless survival rates for patients who underwent this modality with either radical local tumor recurrence represents the real challenge related to MPM.

iMig2012.org • Abstract Book 33 Intrapleural application of chemotherapy and immunotherapy for the SESSION IIIA SURGERY FOR MESOTHELIOMA adjuvant treatment of MPM has been reported. We previously showed September 13, 2012 10:00-11:30 that intrapleural polymeric films loaded with cisplatin were significantly effective in reducing tumor recurrence compared with cisplatin solution IIIA.6: TRIMODALITY THERAPY WITH EXTRAPLEURAL assuring higher and more prolonged plasmatic drug concentrations without PNEUMONECTOMY, RADIATION THERAPY, AND CHEMOTHERAPY increasing toxicity. This study aims to investigate the pharmacokinetic FOR MALIGNANT PLEURAL MESOTHELIOMA profile and tolerability of intrapleural polymeric films containing cisplatin for MPM in an ovine model. Kazunori Okabe1, Eisuke Matsuda1, Hiroyuki Tao1, Tatsuro Hayashi1, Tosiki Tanaka1, Humiho Sano1, Akihiro Takahagi1, Keisuke Aoe2, Koutaro Methods: Hyaluronate films loaded with cisplatin (100mg/m2) previously Taguchi3 characterized were used for the local delivery of anticancer drug. Female 1Thoracic Surgery, Yamaguchi Ube Medical Center, Ube/yamaguchi/ sardinian sheep weighing 40-50kg were chosen for in vivo experiments. JAPAN, 2Medical Oncology, Yamaguchi Ube Medical Center, Ube/ After general anesthesia the sheep were placed in a right lateral decubitus: yamaguchi/JAPAN, 3Radiology, Yamaguchi Ube Medical Center, Ube/ a left pneumonectomy was carried out through a lateral thoracotomy. yamaguchi/JAPAN Thereafter, the adjuvant treatment was randomly administered: september 2012 13, intravenous cisplatin, intrapleural cisplatin, intrapleural hyaluronate- Malignant pleural mesothelioma (MPM) is a serious disease, and a cisplatin. Controls (pneumonectomy alone) were used for comparison. treatment strategy for MPM has not yet been established. We report our Blood samples were taken as scheduled. The animals were euthanatized on experience of trimodality therapy with extrapleural pneumonectomy (EPP), postoperative-day 7: serum, parietal, diaphragmatic pleura, pericardium, radiation therapy, and chemotherapy for MPM. kidneys and liver were considered for analysis. Primary endpoint was plasmatic cisplatin concentration evaluated by IPC-mass spectrometry. Thirty-one EPP were completed in our hospital between June 2006 and Secondary endpoints were treatment-related toxicity and tissue drug January 2012. This number is one of the biggest in Japan within the period, concentration. Data are given as mean. ANOVA was applied for statistical because the Japanese medical system let MPM patients disperse to many analysis. The study was approved by the local veterinary committee. hospitals. Among the thirty-one EPP, twenty-seven consecutive EPP for MPM which were performed by the first author’s thoracic surgery team Results: Three animals per group were treated so far. Mean operation time were reviewed retrospectively. We have instituted a trimodality therapy was 88 minutes (range, 70-127). After 30’ from intravenous administration, protocol consisting of EPP, adjuvant 45 Gy hemithoracic radiation therapy, plasmatic drug concentration was significantly higher (3148ng/ml) than and adjuvant cisplatin-based chemotherapy. Twenty-two patients have intrapleural cisplatin solution (772ng/ml, p=0.007) and intrapleural been treated with this protocol. Five patients were given induction hyaluronate-cisplatin (163ng/ml, p=0.002). At seven days, plasmatic chemotherapy with CDDP and PEM, and referred to us. They underwent cisplatin concentration was much higher (3359ng/ml) after intrapleural EPP and adjuvant radiation therapy. Age, gender, left or right, type of MPM, hyaluronate-cisplatin in comparison to intrapleural and intravenous time of EPP, blood transfusion during EPP, perioperative complication, cisplatin solution [1691ng/ml (p=0.051) and 1264ng/ml (p=0.027), p-TNM and p-Stage, radiation therapy, chemotherapy, and prognosis were respectively] reflecting the controlled drug release from hyaluronate films examined. Overall survival was calculated using the Kaplan-Meier method. (Figure 1). No haematological toxicity was observed. On postoperative-day Survival differences were analyzed with the log-rank test. 7, creatinine levels was significantly higher (p=0.033) after intravenous administration (26.6mg/dl) in comparison to intrapleural hyaluronate- The median age at EPP was 61 years old (44-74). There were 21 males and cisplatin (6.5mg/dl) and controls (1.7mg/dl, p=0.014). Animals treated with 6 females. The right side was affected in 14 and the left side in 13. The intrapleural cisplatin had creatinine levels much higher (22,7mg/dl) than epithelioid type was present in 17, with biphasic type in 6, sarcomatous intrapleural hyaluronate-cisplatin but the difference was not statistically type in 2, and special type in 2. The median EPP time was 7 hours 40 significant (p=0.092). Severe degeneration in tubular cells and glomerular minutes (5 hours 52 minutes – 10 hours 15 minutes). No blood transfusion congestion was microscopically found after cisplatin solution, while a mild during EPP was needed in 12 cases (44%). Mortality involved one patient injury was present after hyaluronate-cisplatin. (3.7%), who died on post-operative day 14 due to the acute aggravation of interstitial pneumonia. Eleven patients (41%) had perioperative international mesothelioma interest group complications. Atrial fibrillation was the most common morbidity, and developed in seven patients (26%). The IMIG pathological TNM was T4 (peritoneal cavity) in 1, T3 in 13, T2 in 6, T1b in 5, N2 in 11, and N0 in 16. The IMIG pathological stage was stage IV in 1, stage III in 17, stage II in 4, and stage Ib in 5. Adjuvant 45 Gy hemithoracic radiation therapy was completed in 23 patients (85%). Six patients (22%) could not undergo adjuvant chemotherapy. Seventeen patients (63%) underwent trimodality therapy. However, seven patients (26%) could not undergo it, and three patients (11%) are currently waiting for adjuvant chemotherapy. The three- year survival, two-year survival, and median survival of all twenty-even patients were 27%, 36%, and 13 months, respectively. The three-year survival, two-year survival, and median survival of seventeen patients who underwent trimodality therapy were 37%, 49%, and 23 months, respectively. The median survival of seven patients who could not undergo trimodality therapy was 6 months. Survival of the patients with trimodality therapy was significantly better than the patients without trimodality therapy.

Trimodality therapy with EPP, radiation therapy, and chemotherapy for MPM is feasible. However, the prognosis of MPM patients should be quickly and markedly improved.

Disclosure: No significant relationships.

Conclusion: Preliminary data showed that intrapleural polymeric films containing cisplatin assured higher plasmatic drug concentration than cisplatin solution without increasing systemic toxicity after seven days.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 34 Session IIIB Apoptosis and Signal Transduction September 13, 2012 10:00-11:30

SESSION IIIB APOPTOSIS AND SIGNAL TRANSDUCTION SESSION IIIB APOPTOSIS AND SIGNAL TRANSDUCTION September 13, 2012 10:00-11:30 September 13, 2012 10:00-11:30 september 2012 13, IIIB.2: MACROPHAGES CAN BE MANIPULATED TO ENHANCE THE IIIB.3: MANIPULATING THE NOXA/BIM AXIS UNDERMINES THE APOPTOTIC RESPONSE TO CHEMOTHERAPY IN MESOTHELIOMA MULTICELLULAR RESISTANCE OF 3D TUMOR SPHEROIDS

Sailaja Battula1, Collin Blakely1, Dario Barbone1, David J. Sugarbaker2, Dario Barbone1, Sailaja Battula1, Hyun-kyung Lee1, David J. Sugarbaker2, Raphael Bueno2, Lisa M. Coussens3, Courtney Broaddus4 Raphael Bueno2, Dean Fennell3, Courtney Broaddus4 1University Of California San Francisco, San Francisco/CA/UNITED STATES 1University Of California San Francisco, San Francisco/CA/UNITED STATES OF AMERICA, 2Division Of Thoracic Surgery, Brigham And Women’s OF AMERICA, 2Division Of Thoracic Surgery, Brigham And Women’s Hospital, Boston/MA/UNITED STATES OF AMERICA, 3Cell & Developmental Hospital, Boston/MA/UNITED STATES OF AMERICA, 3Mrc Toxicology Unit, Biology, Oregon Health And Sciences University, Portland/UNITED STATES University Of Leicester, Leicester/UNITED KINGDOM,4Division Of Pulmonary OF AMERICA, 4Division Of Pulmonary And Critical Care Medicine San And Critical Care Medicine San Francisco General Hospital, University Of Francisco General Hospital, University Of California, San Francisco, San California, San Francisco, San Francisco/CA/UNITED STATES OF AMERICA Francisco/CA/UNITED STATES OF AMERICA Background: When grown as 3D multicellular spheroids, mesothelioma Background: Macrophages within the solid tumor microenvironment cells acquire multicellular resistance to apoptosis. Prior work in our (TME) contribute to tumor chemoresistance by several mechanisms. laboratory has suggested that this resistance arises from alterations in We have found that macrophages constitute a significant percentage of the Bcl-2 family of apoptotic proteins. One mechanism for multicellular the infiltrating leukocyte population in human mesothelioma, a highly resistance to chemotherapy is the lack of up-regulation of the pro- chemoresistant tumor. We investigated whether therapeutic strategies apoptotic sensitizer Noxa which acts by displacing pro-apoptotic Bim from either to reprogram the macrophage phenotype from a Th2-type (pro- its anti-apoptotic buffering proteins allowing Bim to induce apoptosis. tumor) towards a Th1-type (anti-tumor) phenotype or instead to reduce the Interestingly, from prior work, we have found that spheroids contain number of macrophages in mesotheliomas by blockade of the predominant elevated Bim and thus would be sensitive to displacement by Noxa. This macrophage survival pathway, e.g. colony stimulating factor 1 receptor pro-apoptotic potential of the spheroids, termed apoptotic ‘priming’, could (CSF1R), would alter chemoresponsiveness of mesotheliomas. be exploited by therapeutic strategies designed to displace Bim, such as by restoration of Noxa upregulation or by blockers of the anti-apoptotic Methods: To address this, we used two 3D organotypic spheroid models of proteins buffering Bim. When grown as 3D multicellular spheroids, mesothelioma: 1) multicellular spheroids (MCS), with human mesothelioma mesothelioma cells acquire multicellular resistance to apoptosis. Prior work cells grown alone or co-cultured with macrophages derived from in our laboratory has suggested that this resistance arises from alterations peripheral blood monocytes, and 2) primary tumor fragment spheroids in the Bcl-2 family of proteins. One mechanism for multicellular resistance

(TFS), which are small fragments of tumor generated from resected may be a lack of up-regulation of the pro-apoptotic sensitizer Noxa which international mesothelioma interest group human mesothelioma tumor. We also used an in vivo murine model of acts by displacing pro-apoptotic Bim from its anti-apoptotic buffering syngeneic mesothelioma tumor. MCS or TFS were incubated with Th1-type proteins allowing it to induce apoptosis. Interestingly, from prior work, (LPS & IFNg) or Th2-type (IL-4 &IL-13) cytokines, followed by exposure we have found that spheroids contain elevated Bim and thus sensitive to standard-of-care chemotherapy, carboplatin plus pemetrexed. TFS to displacement by Noxa. The pro-apoptotic potential of the spheroids, and mice with syngeneic orthotopic mesothelioma were treated with termed apoptotic ‘priming’, could be exploited by therapeutic strategies carboplatin plus pemetrexed with and without a small molecule inhibitor of designed to displace Bim, such as by restoration of Noxa upregulation or by CSF1R, e.g., GW2580. blockers of the anti-apoptotic proteins buffering Bim.

Results: In both spheroid models, Th1-type macrophage programming Methods: We used several strategies to manipulate Noxa and/or Bim in the significantly increased the apoptotic response of mesothelioma resistant multicellular spheroids. The action of Noxa was replaced by use cells to chemotherapy. Moreover, in the two spheroid models, of a permeable Noxa peptide (R8-Noxa) and Noxa protein was increased when CSF1R signaling in macrophages was inhibited by incubation by use of the histone deacetylase inhibitor, vorinostat/SAHA. Bim was with GW2850, chemoresponsiveness was significantly increased as displaced by the BH3-mimetic, ABT-737, or was increased by the use of evidenced by an increased presence of apoptotic tumor cells. Enhanced SAHA. siRNA was used to knockdown Noxa or Bim to confirm its role in the chemoresponsiveness of mesothelioma tumor cells was found to be apoptotic response to these manipulations. Strategies were also tested in dependent on the presence of macrophages because incubation of tumor fragment spheroids derived from human mesothelioma tumor. tumor cells alone with Th1-type cytokines or with GW2850 was without effect. On the other hand, exposure of macrophages alone to GW2850 in Results: We found that the multicellular resistance of the 3D multicellular vitro resulted in a 50% decrease in macrophage viability. Finally, treatment spheroids could be significantly reduced either by restoring Noxa levels of tumor-bearing mice with GW2580 in combination with carboplatin plus using the permeable Noxa peptide or by releasing Bim via inhibition of the pemetrexed chemotherapy resulted in a significant decrease in tumor anti-apoptotic Bcl-2 proteins using the small molecule inhibitor, ABT-737. burden and an increase in tumor cell apoptosis as compared to treatment Moreover, multicellular resistance was also significantly reduced by use of of mice with chemotherapy or GW2580 alone. SAHA, which upregulated both Noxa and Bim expression. Knockdown of Noxa blocked the effect of SAHA and knockdown of Bim blocked the effect Conclusion: Based on these data, we propose that manipulating the of the Noxa peptide, of ABT-737 and of SAHA showing the importance of macrophage within the mesothelioma TME may be a promising therapeutic each protein in the apoptotic response to these agents. These strategies approach. increased the apoptotic chemoresponsiveness of mesothelioma cells within human tumor fragment spheroids as well. Disclosure: No significant relationships. Conclusion: Using clinically-relevant 3D models, we conclude that

iMig2012.org • Abstract Book 35 manipulation of the core apoptotic repertoire in order to release Bim Pathology, Tokyo Women’s Medical University, Yachiyo/JAPAN may improve the chemosensitivity of mesothelioma. We wish to thank the following for grant support: TRDRP fellowship to DB (18FT-0120), an Ireland/ Background: Majority of malignant mesothelioma is defective of the NCI Consortium grant to VCB, DAF and DB (CDV/3679/07) and a Department of INK4A/ARF locus that contains the p14ARF and p16INK4A genes but Defense Mesothelioma Program grant to VCB (PR080717). possesses the wild-type p53 gene. The characteristic genetic defect leads to inactivation of the p53-mediated pathways together with cell cycle Disclosure: No significant relationships. progression through phosphorylated pRb since p14ARF blocks Mdm2- mediated p53 degradation and p16INK4A inhibits cyclin-dependent kinases. These properties can contribute the uncontrolled cell proliferation with enhanced resistance to apoptotic stimuli. SESSION IIIB APOPTOSIS AND SIGNAL TRANSDUCTION September 13, 2012 10:00-11:30 Methods: We thereby examined a possible therapeutic strategy by restoring the p53 functions with adenoviruses expressing wild-type p53 IIIB.4: THE HSP90 INHIBITOR GANETESPIB REQUIRES THE BID (Ad-p53) and Ad defective of p53-binding E1B55kDa molecules (Ad- DRIVEN MITOCHONDRIAL PATHWAY EXECUTE APOPTOTIC CELL delE1B), and further investigated combinatory cytotoxic effects with september 2012 13, DEATH IN MESOTHELIOMA cisplatin (CDDP) and pemetrexed (PEM). Ad-delE1B can replicate and produce the progenies in target cells whereas Ad-p53 are replication- Sara Busacca1, Astero Klabatsa2, Michael Sheaff3, Dean Fennell1 incompetent. Cytotoxicity and cell cycle were assessed with colorimetric 1University Of Leicester, Leicester/UNITED KINGDOM, 2Guy’s Hospital, assay and flow cytometry, respectively. Expression levels of p53-associated Guy’s And St Thomas’ Nhs Foundation Trust/UNITED KINGDOM, 3Barts And and apoptosis-linked proteins were also examined with Ad expressing beta- The London Nhs Trust/UNITED KINGDOM galactosidase gene as a control. The anti-tumor activity was tested in an orthotopic animal model by injecting mesothelioma in the pleural cavity. Background: Mesothelioma is a highly antiapoptotic cancer. This phenotype may be in part related to multiple constitutively activated Results: Transduction of human mesothelioma cells with Ad-p53 or growth factor signals. HSP90 inhibition may be a clinically useful strategy Ad-delE1B up-regulated p53 expression levels and induced the p53 for suppressing multiple growth factor signals via the PI3K/AKT/mTOR and phosphorylation at Ser 15 and 46 residues, activation markers of p53, RAS/RAF/MEK pathways in mesothelioma. A phase 1/II trial is currently in and subsequently increased expression of p21 and Mdm2, the p53 target development in the UK, called MESO2 (NCT01590160). molecules. An augmented expression of p21, an inhibitor of cyclin- dependent kinases, dephosphorylated pRb and induced cell cycle arrest Methods: Mouse embryonic fibroblasts (MEFs) with either wild type at G1 phase. The Ad-transduced cells activated primarily the extrinsic (WT) of double knockout (DKO) of BAX/BAK were treated with the HSP90 apoptotic pathways accompanied by enhanced Fas and TRAIL receptor inhibitor, ganetespib, and IC50 values determined. RNAi was used in two expressions. The transduction subsequently showed cleavage of capase-8 mesothelioma cell lines (MSTO-211H and NCI-H2052) to silence BAX/BAK. but less significantly caspase-9, and increased sub-G1 cell populations. A Apoptosis was analysed by poly ADP-ribose polymerase (PARP) cleavage combinatory use of Ad-p53 or Ad-delE1B with CDDP or PEM, the first- and caspase 9 (C9) activation. Focused RNAi array targeting of BH3 only line agents for mesothelioma, showed synergistic cytotoxic effects. We proteins (which activate BAX/BAK or antagonise prosurvival BCL2 proteins) also demonstrated the anti-tumor effects of Ad-p53 and Ad-delE1B in was conducted to delineate critical death activators. an orthotopic animal model. Intrapleural injection of Ad-p53 or Ad- delE1B decreased the tumor weight of human mesothelioma developed Results: Double knockout or RNAi silencing rescued cells from ganetespib- in the pleural cavity of immunocompromised mice and further achieved mediated apoptosis as evidenced by reduced PARP/C9 cleavage, combinatory effects with intraperitoneal CDDP administration. Interestingly mitochondrial depolarisation and sub-G1 analysis and increased viability. Ad-delE1B55-infected cells displayed a hyperploid state at the cell cycle Systematic functional assessment of BH3 proteins identified BID silencing analysis and showed enlarged nuclear configurations followed by pyknotic as being sufficient to rescue cells. We have found that loss of both BAX nuclear changes that were positive for a TUNEL assay. and BAK is observed in 37% of primary mesotheliomas. Loss of BID in international mesothelioma interest group mesothelioma has been reported to be 38% Conclusion: Intrapleural injection of Ad-p53 or Ad-delE1B in combination with systemic administration of the first-line agents is a feasible Conclusion: Inhibition of HSP90 requires the mitochondrial pathway therapeutic strategy for mesothelioma by inducing apoptotic cell death. to induce cell death. Our data suggest that loss of BAX/BAK or BID may be sufficient to induce resistance, and could be putative biomarkers of Disclosure: No significant relationships. sensitivity to ganetespib, and provides a hypothesis for clinical correlation in trial NCT01590160.

Disclosure: No significant relationships. SESSION IIIB APOPTOSIS AND SIGNAL TRANSDUCTION September 13, 2012 10:00-11:30

IIIB.6: DEATH EFFECTORS – DISSECTING MECHANISMS OF SESSION IIIB APOPTOSIS AND SIGNAL TRANSDUCTION APOPTOSIS RESISTANCE September 13, 2012 10:00-11:30 Marion MacFarlane IIIB.5: REACTIVATION OF P53-MEDIATED PATHWAYS INDUCES Toxicology Unit, Mrc, Leicester/UNITED KINGDOM APOPTOSIS IN MESOTHELIOMA WITH WILD-TYPE P53 GENE AND PRODUCES COMBINATORY SYNERGISTIC EFFECTS WITH ANTI- Acquisition of resistance to apoptosis is a defining feature of tumour cells, CANCER AGENTS thus strategies that bypass nodes of apoptotic resistance may be used to trigger cell death selectively in tumour cells. Therapeutic strategies Masatoshi Tagawa1, Quanhai Li1, Kiyoko Kawamura1, Masato Shingyoji2, to manipulate apoptosis have immense potential as they could provide Yuji Tada3, Yuichi Takiguchi4, Koichiro Tatsumi3, Hideaki Shimada5, viable alternatives to currently available therapeutic options associated Kenzo Hiroshima6 with high toxicity and numerous adverse side effects. While the profound 1Division Of Pathology And Cell Therapy, Chiba Cancer Center Research resistance of mesothelioma to cytotoxic agents is well documented, and Institite, Chiba/JAPAN, 2Department Of Thoracic Disease, Chiba Cancer is reportedly due to inhibition of cell death, the identification of which cell Center, Chiba/JAPAN, 3Department Of Respirology, Chiba University School death pathways are deregulated in this disease has not been systematically Of Medicine, Chiba/JAPAN, 4Department Of Medical Oncology, Chiba examined. Dissecting the molecular mechanisms that determine death or University School Of Medicine, Chiba/JAPAN, 5Department Of Surgery, survival of tumour cells following exposure to a panel of agents that target Toho University School Of Medicine, Tokyo/JAPAN, 6Department Of specific nodes of apoptosis resistance is therefore of key importance.

iMig2012.org • Abstract Book 36 To address this, we have employed both mechanistic and translational- based approaches with the aim of identifying which cell death pathways are deregulated in mesothelioma by examining the expression of key cell death pathway regulators in different subtypes of human mesothelioma cell lines, normal untransformed mesothelial cells and mesothelioma tissue obtained from patients. In addition, we have compared the sensitivity of normal mesothelial cells and mesothelioma cell lines to a range of cytotoxic agents, including DNA damaging agents and novel agents that target specific nodes of apoptosis resistance. Importantly, based on initial profiling of malignant mesothelioma tumour samples from patients, several potentially viable drug targets for modulating tumour cell death/ keyregulators of apoptosis signalling pathways, will be highlighted (e.g. Inhibitors of BCL-2, TRAIL-Receptor agonists, and Inhibitors of glycolysis such as 2-deoxyglucose). This work was supported by the Medical Research Council (UK) september 2012 13,

Disclosure: No significant relationships. international mesothelioma interest group

iMig2012.org • Abstract Book 37 Session IIIC Mesothelioma Epidemiology September 13, 2012 10:00-11:30

SESSION IIIC mESOTHELIOMA EPIDEMIOLOGY SESSION IIIC mESOTHELIOMA EPIDEMIOLOGY September 13, 2012 10:00-11:30 September 13, 2012 10:00-11:30 september 2012 13, IIIC.2: EXPOSURE TO ASBESTOS IN HOME RENOVATION AND IIIC.3: MALIGNANT MESOTHELIOMA IN ITALY: INCIDENCE, SUBSEQUENT RATES OF MALIGNANT MESOTHELIOMA IN MODALITIES OF ASBESTOS EXPOSURE AND OCCUPATIONS WESTERN AUSTRALIA INVOLVED FROM THE ITALIAN NATIONAL REGISTER.

Nicholas De Klerk1, Alison Reid2, Nola Olsen2, Peter Franklin2, Alessandro Marinaccio1, Marina Verardo2, Dario Mirabelli3, Valerio Arthur W. Musk3 Gennaro4, Carolina Mensi5, Gert Schallenberg6, Enzo Merler7, Renata 1Institute For Child Health Research, University Of Western Australia, De Zotti8, Antonio Romanelli9, Elisabetta Chellini10, Cristiana Pascucci11, Subiaco/WA/AUSTRALIA, 2School Of Population Health, University Of Daniela D’Alò12, Francesco Forastiere13, Simona Menegozzo14, Luana Western Australia, Crawley/WA/AUSTRALIA, 3Respiratory Medicine, Sir Trafficante15, Marina Musti16, Gabriella Cauzillo17, Attilio Leotta18, Charles Gairdner Hospital, Nedlands/WA/AUSTRALIA Rosario Tumino19, Massimo Melis20 1Inail, Research Area, Roma/ITALY, 2Cor Valle D’Aosta, Aosta/ITALY, 3Cor Background: Worldwide rates of malignant mesothelioma (MM) have been Piemonte, Torino/ITALY, 4Cor Liguria, Genova/ITALY, 5Cor Lombardia, driven largely by occupational exposure to asbestos and the burden of Milano/ITALY, 6Cor Pa Trento, Bolzano/ITALY, 7Cor Veneto, Padova/ disease from this exposure will continue for many years. Recent concerns ITALY, 8Cor Friuli-Venezia Giulia, Trieste/ITALY, 9Cor Emilia-Romagna, about additional future incidence of MM have been raised concerning Reggio Emilia/ITALY, 10Cor Toscana, Firenze/ITALY, 11Cor Marche, Ancona/ exposure to asbestos arising from home renovations involving existing ITALY, 12Cor Umbria, Perugia/ITALY, 13Cor Lazio, Lazio (roma)/ITALY, 14Cor asbestos-containing materials (ACM) (Olsen et al, Med J Aust 2011;195:271- Campania, Napoli/ITALY, 15Cor Abbruzzo, Tocco Di Casauria/ITALY, 16Cor 274). Those results were, however, based on numbers of cases, not rates Puglia, Bari/ITALY, 17Cor Basilicata, Potenza/ITALY, 18Cor Calabria, Lamezia in this population. This study aimed to estimate rates of MM among people Terme/ITALY, 19Cor Sicilia, Ragusa/ITALY, 20Cor Sardegna, Cagliari/ITALY who had received their exposure from renovations in homes with ACM in place. Background: Due to the large scale use of asbestos (more than 3.5 million tons produced or imported until its definitive banning in 1992), a specific Methods: The Western Australian (WA) Mesothelioma Register has national surveillance system of mesothelioma incident cases is active in recorded exposure and disease details for all cases of MM occurring in Italy, with direct and individual anamnestic etiological investigation. WA since the first case in 1961. All cases attributable to home renovation exposure were collated and their characteristics have been reported, Methods: In each Italian Region there is a epidemiological center to indicating a large rise in numbers over the last 10-15 years. A computer– actively search and collect every mesothelioma incident case from health assisted telephone survey in 2008 interviewed 2800 people in Australia care institutions that diagnose and treat them. These include pathology

(700 in WA) about previous occupational and environmental exposure to and histology units, lung disease and chest surgery wards. Occupational international mesothelioma interest group asbestos. The prevalence of home exposure, without any occupational history, lifestyle habits and areas of residence for each case are obtained exposure, for different age groups and for different times from first by interviewing people directly or, if they are not available, someone close exposure and duration of exposure were estimated from WA responders. to them (indirect interview) who can provide information on the case’s These proportions were then applied to the total population obtained from work and life history. Exposure is classified by an industrial hygienist using the Australian Bureau of Statistics and used as denominators to estimate the standard grid in accordance with national guidelines and a standard rates. questionnaire, administered by a trained interviewer.

Results: The survey indicated that around 15% of the population had Results: In the period between 1993 and 2008, a case-list of 15,845 MM been exposed to asbestos arising from home renovation that, based on was recorded by the Italian National Register (ReNaM) and the modalities reported activities, was equivalent to that experienced by cases of MM. of exposure to asbestos fibres have been investigated for 12,065 of them. Estimated rates increased with both time since first exposure and duration Age at diagnosis is lower than 55 years for 9.4% of cases and the mean of exposure. Averaging over all durations of exposure of more than 7 days, is 69.2 years at diagnosis. Gender ratio (M/F) is 2.5. The anatomical and time since first exposure greater than 10 years, MM rates ranged from sites of the disease is the pleura for 94% of cases; peritoneum for 6.4%. 2 per million person-years under age 40 to 80 per million over age 75, Standardized incidence rates are 3.84 (per 100,000 inhabitants) for more than 100 times less than rates after exposure to crocidolite at the men and 1.45 for women, with a wide regional variability. Occupational Wittenoom asbestos mine, but greater than background rates. asbestos exposure was in 69.3% of interviewed subjects cases, while 4.4% was due to cohabitation with someone (generally, the husband) Conclusion: Although increased, absolute risks from home renovation occupationally exposed, 4.3% by environmental exposure from living near exposure are not high. As well as quantifying the risk from this particular a contamination source and 1.6% during a leisure activity. In the exposed kind of exposure, these data will also enable us in future work to assign workers, the median year of first exposure was 1957, and mean latency risks to other groups of exposed people, both occupational and non- was 46 years. occupational, and to make realistic projections of future incidence of MM in these groups. Conclusion: The analysis of exposures focuses on a large variety of economic sectors involved and not only for those traditionally signaled Disclosure: No significant relationships. as ‘‘at risk’’ (like asbestos-cement industry, shipbuilding and repair and railway carriages maintenance) and an increasing trend for the building construction sector. The systematic mesothelioma surveillance system is relevant for the prevention of the disease and for supporting an efficient compensation system. Our data illustrate the importance of documentation and dissemination of all asbestos exposure. The existing

iMig2012.org • Abstract Book 38 experience on asbestos effect must to be transferred to developing SESSION IIIC mESOTHELIOMA EPIDEMIOLOGY countries where asbesto use is spreading. September 13, 2012 10:00-11:30

Disclosure: No significant relationships. IIIC.5: MESOTHELIOMAS SHOWN DIFFERENCES WHEN OCCURRING AMONG EXPOSED OR UNEXPOSED AND UNLIKELY EXPOSED TO ASBESTOS

SESSION IIIC mESOTHELIOMA EPIDEMIOLOGY Enzo Merler, Vittoria Bressan, Paolo Girardi, Regional Group On se SEptember 13, 2012 10:00-11:30 Malignant Mesothelioma Regional Group On Malignant Mesothelioma Occupational Health Department, Local Health Authority, Venetian IIIC.4: MESOTHELIOMA INCIDENCE IN AN ASBESTOS-EXPOSED Mesothelioma Register, Padova/ITALY COHORT IN SOUTH AFRICA: PREDICTING CASES INTO THE FUTURE Background: With the aim to evaluate the asbestos exposure, we attempted to interview and collect information on all confirmed Gill Nelson1, Benn Sartorius1, Jill Murray2, Tobias Chirwa1, Markus Heitz3, mesothelioma (MM) cases occurred between 1987 and 2011 in the Veneto september 2012 13, Jim Tewaternaude4 Region, Italy (4.5 million inhabitants), in the framework of a population- 1Epidemiology And Biostatistics, School Of Public Health, University based mesothelioma Registry, part of the National Mesothelioma Of The Witwatersrand, Johannesburg/SOUTH AFRICA, 2Pathology, Registry. Following predefined national guidelines, asbestos exposure National Institute For Occupational Health, Nhls, Johannesburg/ was assigned to each investigated case by circumstance (occupational, SOUTH AFRICA, 3Fmh Innnere Medizin+ Pneumologie, Zurich/ familial, environmental) and probability (definite, probable, possible, SWITZERLAND,4School Of Public Health And Family Medicine, University Of unlikely, unknown). Severity of asbestos exposure is assumed to decrease Cape Town, Cape Town/SOUTH AFRICA from definite occupational exposure through leisure-time and unknown or unlikely. It is reasonable to assume that asbestos exposure influences the Background: South Africa mined and milled all three commercial forms occurrence and distribution by site of MM and other characteristics. of asbestos (crocidolite, amosite and chrysotile) for more than 100 years, with production peaking at around 350 000 tons in the 1970s. Miners, Methods: We evaluate in a case-case comparison differences among MM millers and community members were exposed to high levels of fibres with asbestos exposure (certain, probable and possible occupational) occupationally and environmentally. There is currently worldwide interest (n. 1140), and environmental and domestic exposure (n. 214) (“exposed in mesothelioma trends and high rates have been reported in South cases”) vs MM without (n. 18) and not classifiable for asbestos exposure (n. Africa. Asbestos was mined in South Africa until 2002, and only banned 211) (“unexposed cases”). The latter may be considered as MM occurred in 2009, so rates are not expected to decline soon. South African laws among subjects without an easily detectable asbestos exposure. The allow for the financial compensation of occupationally acquired asbestos- Male:Female ratio (M:F) was calculated and tested in relation to exposure related diseases but no compensation is provided to those exposed and site (pleural vs. peritoneal). The association between site and exposure environmentally. The Asbestos Relief and Kgalagadi Relief Trusts provide was estimated using Odds Ratios (OR and 95% Confidence Intervals, CI) by additional compensation for diseased persons who worked at or lived in logistic regression models. the vicinity of certain asbestos mines in South Africa. Since 2003, almost 15 000 potential claimants have registered with the Trusts; some 4 800 Results: The M:F among “unexposed cases” approximated unity. The had compensable claims, including 390 with malignant mesothelioma. ratio was strongly increased among occupationally “exposed” (7.68), the It is important for the Trusts to estimate the number of future cases as highest value observed for pleural MM severely exposed (12.86). Among accurately as possible, to equitably allocate remaining funds. The objective non-occupationally exposed the M:F was below unity. M:F by MM site and of this study was to develop robust mathematical and statistical models to asbestos exposure. accurately predict the number of cases of mesothelioma likely to present over the next 10 to 15 years. international mesothelioma interest group Certain, Possible Environ- Without. p-value Methods: Demographic and relevant asbestos exposure data (such probable occu- mental, non as job type and asbestos exposure levels), captured by the Trusts, occupational pational domestic classifiable were used to construct three prediction models which were based on (1) (2) (3) (4) classical, deterministic and Bayesian statistical methods developed in other countries. The outcome was death due to mesothelioma. A M:F M:F M:F M:F number of predictor variables, such as date of birth, date of death, age Pleural 12.86 2.33 0.36 1.08 <0.001 at first exposure, age at last exposure, job exposure category, duration of exposure, year(s) of exposure, and year-specific asbestos fibre Peritoneal 9.2 1.4 0.19 0.82 <0.001 measurements, were used to estimate model parameters. All three models were assessed in terms of their goodness of fit and predictive capabilities. The risk of a pleural MM (vs peritoneal) increases by severity; exposure 1: The model with the best fit was selected as the final model for prediction OR 3.28 (1.93-5.57); exposure 2: OR 2.95 (1.44-6.05); exposure 3: OR 1.94 purposes. The data were managed and analysed in STATA version 12.0 SE. (1.07-3.54). The risk of a pleural MM (vs. peritoneal) increases by 4% at any additional year of age at diagnosis: OR: 1.04 (1.02-1.06). Results: From 2003 to 2011, 390 cases of mesothelioma (2.6%) were diagnosed in the cohort of 14 738 individuals. Their mean age was 59.2 Conclusion: Our study confirmes that asbestos increases MM according to years (SD 10.5) and the mean latency period since first exposure was 30.2 severity of exposure with a different exposure-response relation by site. All years (SD 9.1). Exposure history data show a peak exposure in 1982 (in MM pleural cases that our criteria define as exposed should be considered around 30% or 4 370 of the cohort). Preliminary estimates suggest that to be caused by asbestos exposure and therefore eligible for compensation. mesothelioma incidence will peak in 2016. Disclosure: No significant relationships. Conclusion: With the incidence yet to peak, allocation of the Trusts’ remaining funds remains critical. More advanced modelling techniques will be applied to confirm these predictions. Updating the cohort vital status, using data from Statistics South Africa, will improve the predictive power of the models.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 39 SESSION IIIC mESOTHELIOMA EPIDEMIOLOGY September 13, 2012 10:00-11:30

IIIC.6: MESOTHELIOMA IN ASIA

Ken Takahashi Department Of Environmental Epidemiology, University Of Occupational And Environmental Health/JAPAN

Despite the universal consensus on the medical entity of mesothelioma, many countries, mostly of economically developing status, do not recognize or report the disease. This could be due to a variety of reasons: (a) historical use and exposure to asbestos may have been nil or negligible; (b) historical exposure may have occurred but started only recently, so the latency time has not yet reached saturation. In other words, the disease is september 2012 13, only waiting to surface; (c) the clinical, pathological and social resources required for diagnosis are not yet available. In other words, the disease is there, but detection is compromised by the lack of technology and resources. A less likely but another possible reason is that (d) the disease is diagnosed but not officially reported (intentionally or otherwise). In Asia (its definition differs even between the United Nations Statistics Division and the World Health Organization, but, here, in the general sense), most countries are of economically developing status. Most, if not all, of Asian developing countries do not officially diagnose/report mesothelioma. According to the WHO Mortality Database, since the disease entity of mesothelioma appeared therein, about 12,000+ mesothelioma deaths* have been reported cumulatively in Asian countries. The vast majority of this is reported by only a few developed countries, i.e., Japan, Korea and Singapore. In contrast, Asian developing countries report almost no mesothelioma, for at least one of the aforementioned reasons. I further speculate the breakdown of reasons to be roughly, (a) minimal, (b) substantial, (c) substantial, and (d) unknown. It should also be noted that the country status of (b) and (c) is often intertwined. Thus if we account for the historical situation on asbestos use in Asian developing countries, we can expect that the latency time for mesothelioma is approaching saturation in many of these countries. And even when the latency time is reached, the medical and social infrastructure to diagnose, treat and compensate mesothelioma in developing countries is grossly inadequate. Consequently, Asia is in urgent need of attention and action for prevention of mesothelioma, at all respective levels of primary (prevention of exposure), secondary (detection of disease) and tertiary (treatment and compensation). In 2007, the 60th World Health Assembly endorsed a global plan of action (GPA) on workers’ health 2008–2017, and made reference to a global campaign for the elimination of asbestos- international mesothelioma interest group related diseases (ARDs), including mesothelioma. Priority 1.3 of GPA 2009–2012 was to “Develop and disseminate evidence-based tools and raise awareness for the elimination of ARDs”, which later developed into Priority 1.2 of GPA 2012–2017, under the “Regional and national programs on occupational cancer, silica and ARDs”. The Asian initiative to eliminate ARDs (the Asian Asbestos Initiative, or AAI), which was embarked upon by the speaker and colleagues in 2008, achieved widespread recognition and became a formal component of the GPA. The fourth international seminar (AAI-4) was successfully organized by Korean colleagues in 2011, and AAI-5 is planned as a joint Korea-Japan endeavor in 2012. The AAI aspires to provide a model for the world that will pave the way for the ultimate elimination of ARDs, including mesothelioma. *This is equivalent to only 13% of the cumulative number of mesothelioma deaths reported in the world during the same period. By contrast, the proportion of global asbestos use attributed to Asia has been steadily increasing over the years from 14% (1920–1970) to 33% (1971–2000) to 64% (2001–2007). This increase has been reflected in the absolute level of per capita use across a wide range of countries.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 40 General Session IV New Directions and Future Studies II September 13, 2012 13:15-14:15

GENERAL SESSION IV NEW DIRECTIONS AND FUTURE STUDIES II September 13, 2012 13:15-14:15 september 2012 13, GSIV.1: NEW DIRECTIONS AND FUTURE STUDIES IN PERITONEAL MESOTHELIOMA

Paul H. Sugarbaker Program In Peritoneal Surface Malignancy, Washington Cancer Institute, Washington, DC/UNITED STATES OF AMERICA

Background: Peritoneal mesothelioma has been regarding in the past as a lethal disease with approximately a one year median survival. New multimodality treatments utilizing cytoreductive surgery and perioperative chemotherapy are being investigated.

Methods: Radiologic studies, histopathologic studies, improvements in surgical technique, and refinements in perioperative and long-term bidirectional chemotherapy have occurred.

Results: Preoperative biopsy can be used to select patients for aggressive surgical management versus palliative systemic chemotherapy. Also, radiologic studies using an interpretative classification of the abdominal and pelvic CT allows the surgeon to select patients who have a high likelihood of complete cytoreduction. At surgery, peritonectomy procedures are essential for complete tumor resections. An important new modality is hyperthermic intraperitoneal chemotherapy. In addition, long-term bidirectional chemotherapy using an intraperitoneal port has become an essential part of management. The regimens for bidirectional intraoperative hyperthermic chemotherapy, early postoperative intraperitoneal chemotherapy, and adjuvant bidirectional chemotherapy will be presented.

international mesothelioma interest group Conclusions: Information from randomized trials is unlikely to benefit in this rare disease. Using pharmacologic studies and clinical refinements, great progress has been made. Uniform staging and comprehensive reports from all centers of excellence for peritoneal mesothelioma are now indicated.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 41 Session IVA Gene Regulation and Mesothelioma Pathogenesis 3 September 13, 2012 14:20-16:00

SESSION IVA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 3 genes lead to hyperactivation of this pathway in malignant mesothelioma September 13, 2012 14:20-16:00 (MM). september 2012 13, IVA.2: GLI FAMILY OF TRANSCRIPTION FACTORS AS A NOVEL Methods: Activation of the pathway in 6 MM cell lines was determined THERAPEUTIC TARGET IN MALIGNANT PLEURAL MESOTHELIOMA by measuring mRNA levels of the downstream effectors GLI1 and GLI2. Exonic sequencing of 13 HH pathway genes was performed on the MM cell Hui Li, Tiffany Cheng, Natalie Lui, Hanh Do, Katty Tseng, David Jablons, lines. All variants found were confirmed by a second independent PCR and Biao He sequencing reaction. In silico analysis using SIFT program was performed Surgery, University Of California, San Fancisco, San Francisco/CA/UNITED to predict the likelihood that an amino-acid substitution would have an STATES OF AMERICA impact on protein function.

Background: Malignant mesothelioma is a highly aggressive tumor with Results: Hyperactivation of the HH pathway was observed in all cell lines. poor prognosis. Current treatment is rarely curative, thus novel meaningful We found PTCH1, SMO and Suppressor of fused (SUFU) mutations in 3 of therapies are urgently needed. Inhibition of Hedgehog signaling at the cell 6 MM cell lines examined. We identified a novel non-synonymous SUFU membrane level in several cancers has shown anti-cancer activity in recent mutation in exon 10. LO68 contained a novel missense mutation in SUFU clinical studies. Evidence of non-canonical Gli activation suggests Gli as a (p.T411M), which resulted from a nucleotide 1232 C>T transition. Deletion more potent therapeutic target. of six exons in the PTCH1 gene was found in JU77, which resulted in loss of one of two extracellular loops implicated in HH ligand binding and the Methods: RT-PCR and immunohistochemistry were performed to intracellular C-terminal domain. We also detected a novel 3-bp insertion analyze Gli1/2 expression in 42 mesothelioma tissue samples. Cultured (69 _ 70insCTG) in exon 1 of SMO, predicting an additional leucine residue mesothelioma cell lines were employed to investigate roles of Gli activation in the signal peptide segment of SMO protein. None of the cell lines had in mesothelioma. A novel small molecule Gli inhibitor (Gli-I) that we mutations in SHH, DHH, IHH, PTCH1, PTCH2, HHIP, KIF7, SUFU, GLI2 recently developed and siRNA were applied to inhibit Gli. MTS assay was and GLI3. In silico characterization of the SUFU mutant by SIFT program conducted to examine cell proliferation after different treatments. MS1 suggested that the p.T411M mutation might alter protein function. xenograft model was used in in vivo study of the Gli-I. Conclusion: In this study, we identified the first novel mutations in PTCH1, Results: 79% mesothelioma specimens had higher Gli1 or Gli2 expression SUFU and SMO associated with MM. In silico data analysis indicate the than that in adjacent normal tissues by RT-PCR, and 72% were Gli1 or Gli2 SUFU mutation may aberrantly activate the HH pathway. Our data points positive with nuclear localization by immunohistochemistry. Inhibition to a possible driving role for PTCH1 and SUFU in the pathogenesis of a of Gli by shRNAs or Gli-I suppressed cell growth and downregulated subgroup of MM and rationalize the exploration of HH pathway inhibitors Gli downstream targets in vitro. Efficacy of Gli-I (IC50: 0.75~13uM) in for MM therapy.

mesothelioma cell lines correlated with their Gli expression levels. A 14-day international mesothelioma interest group i.p. administration of Gli-I significantly suppressed tumor growth in vivo. Disclosure: No significant relationships. Combination of Gli-I and pemetrexed demonstrated synergistic effect in suppression of mesothelioma proliferation in vitro.

Conclusion: Gli activation plays a critical role in mesothelioma. Inhibition SESSION IVA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 3 of Gli function has strong potential to become a novel, clinically effective September 13, 2012 14:20-16:00 approach to treat mesothelioma. IVA.4: HIPPO SIGNALING PATHWAY INACTIVATION IN Disclosure: No significant relationships. MALIGNANT MESOTHELIOMA CELLS

Yoshitaka Sekido, Ichidai Tanaka, Hirotaka Osada, Makiko Fujii Division Of Molecular Oncology, Aichi Cancer Center Research Institute, SESSION IVA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 3 Nagoya/JAPAN September 13, 2012 14:20-16:00 Background: Malignant mesothelioma (MM) is an aggressive neoplasm IVA.3: GENETIC ALTERATIONS IN THE HEDGEHOG SIGNALING associated with asbestos exposure. MM shows frequent mutation of PATHWAY GENES OF HUMAN MALIGNANT MESOTHELIOMA CELL the neurofibromatosis type 2 (NF2) gene, whose protein product, LINES merlin, regulates several important signaling cascades including the Hippo signaling cascade that controls organ size, development and Chuan Bian Lim, Huimin Cheah, Svetlana Baltic, Philip J. Thompson, differentiation.We previously reported that the LATS2 or SAV1 genes, which Cecilia M. Prele, Steven E. Mutsaers encode a component of this cascade, can also be inactivated in a subset of Lung Institute Of Western Australia, Perth/WA/AUSTRALIA mesotheliomas, which leads to a constitutive activation of Yes-associated protein (YAP) transcriptional coactivator. We also found that YAP activation Background: The Hedgehog (HH) pathway plays a critical role during with TEAD transcription factor upregulates multiple gene transcription embryonic development by regulating proliferation, differentiation and including cell cycle-regulating genes such as CCDN1 and FOXM1. tissue patterning. Recent studies have identified important regulatory roles for this pathway in certain cancers with mutations in the HH pathway Methods: Twenty MM cell lines including cell lines established from genes Patched1 (PTCH1) and Smoothened (SMO) leading to pathway Japanese patients were analyzed. Expression profiling of mRNA was hyperactivation in medulloblastoma and basal cell carcinoma. The aim of conducted with Agilent 44K microarray. Western blot analysis, real-time this study was to test the hypothesis that mutations in the HH pathway RT-PCR, transcriptional reporter assay, ChIP analysis, and cell proliferation

iMig2012.org • Abstract Book 42 and soft agar colony formation assays were performed using standard uncommon in mesothelioma. Protein expression of NF2 and INPP4B was techniques. For animal experiments, 7-week-old female nude mice of KSN lost in 70% and 50% of cell lines respectively and it was more frequent in strain were used. the tissue samples. In addition rare loss of NHERF1 and LKB1 was observed. PTEN was universally present at the protein level mirroring findings at the Results: We demonstrated that the connective tissue growth factor (CTGF) DNA level. GDC-0980 was very potent across all 20 mesothelioma cell lines gene is also an important target gene of YAP in mesothelioma cells, and its analyzed with an average IC50 of <0.9uM. GDC-0980 induced apoptosis upregulation is induced by cooperation with the activation of transforming in ~5-18% cells in a selection of four mesothelioma cell lines. Furthermore, growth factor (TGF)-beta signaling. Enhanced CTGF expression caused GDC-0980 reversed the invasive morphology of the mesothelioma cell lines abundant extracellular matrix formation in vivo, and nude mice which suggesting possible effects of PI3K inhibition on cell motility and invasion. were transplanted with a CTGF-knocked down MM cell line showed Additional PI3K pathway inhibitors with different target specificities were prolonged survival compared to its parental cell line. Meanwhile, since investigated that showed marked activity, albeit with potency not as four of 20 MM cell lines had neither mutation of NF2, LATS2, or SAV1, broadly consistent as GDC-0980. we examined whether other molecules involved in the Hippo signaling cascade are altered in the cell lines. Among the components recently Conclusion: Epithelial pleural mesotheliomas are characterized by frequent identified to regulate the Hippo signaling cascade, we found that Ajuba, loss of the INPP4B and NF2 tumor suppressors that have been linked with september 2012 13, a LIM domain-containing protein, was downregulated in six cell lines, PI3K pathway activation. Contrary to prior reports PTEN is not typically lost with 4 cell lines showing no alteration of the other components of the in epithelial pleural mesotheliomas neither at the DNA nor protein level. Hippo pathway. We transduced exogenous Ajuba into MM cell lines and GDC-0980 has very potent in-vitro activity in all mesothelioma cell lines detected the suppression of cell proliferation. Since one of the multiple and induces apoptosis. Since the correlation of INPP4B and NF2 loss with functions of Ajuba is thought to be adherence junction stabilization, our inhibitor sensitivity is not intuitive, additional model systems or samples results suggested that the cell-cell adhesion status might influence MM cell from patients treated with GDC-0980 need to be evaluated. proliferation via Ajuba-Hippo signaling cascade. Disclosure: No significant relationships. Conclusion: CTGF is an important modulator of MM growth and pathology, whose expression is enhanced by TGF-beta signaling activation and Hippo signaling inactivation. Our results indicate that the dysregulation of Merlin-Hippo signaling in MM cells may also be affected not only by the SESSION IVA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 3 inactivation of core components of this cascade but also its other effectors. September 13, 2012 14:20-16:00

Disclosure: No significant relationships. IVA.6: FIBROBLAST GROWTH FACTOR SIGNALING – A NEW TARGET IN MESOTHELIOMA THERAPY?

Karin Schelch1, Mir A. Hoda2, Christine Pirker1, Bahil Ghanim2, SESSION IVA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 3 Thomas Klikovits2, Viktoria Laszlo2, Ulrike Setinek3, Martin Filipits1, September 13, 2012 14:20-16:00 Balazs Dome2, Balazs Hegedus2, Walter Klepetko2, Walter Berger1, Michael Grusch1 IVA.5: FREQUENT LOSS OF INPP4B AND NF2/MERLIN TUMOR 1Department Of Inner Medicine I, Medical University Of Vienna, Insitute SUPPRESSORS IN MALIGNANT PLEURAL MESOTHELIOMA AS Of Cancer Research, Vienna/AUSTRIA, 2Department Of Surgery, Medical NOVEL CANDIDATE MECHANISMS OF PI3K ACTIVATION AND University Of Vienna, Division Of Thoracic Surgery, Vienna/AUSTRIA, 3Otto GDC-0980 SENSITIVITY. Wagner Hospital, Vienna/AUSTRIA

Tanguy Y. Seiwert1, Arun Khattri1, Jeffrey J. Wallin2, Rajani P. Kanteti3, Background: Malignant pleural mesothelioma (MPM) is an aggressive Zhixiang Zuo1, Michaela K. Keck1, Aliya N. Husain4, K A. Edgar2, Carol malignancy characterized by frequent resistance to chemo- and O’Brien2, Hedy L. Kindler1, Mark Lackner2, Lori S. Friedman2, Ravi Salgia1 radiotherapy, poor outcome and limited therapeutic options. Fibroblast international mesothelioma interest group 1Section Of Hematology/Oncology, University Of Chicago, Chicago/IL/ growth factors (FGF) and their receptors (FGFR) contribute to malignant UNITED STATES OF AMERICA, 2Genentech, South San Francisco/UNITED growth in several tumor types including thoracic malignancies while a role STATES OF AMERICA, 3Department Of Medicine, The University Of Chicago, in MPM remains largely undefined. Therefore, the aim of the present study Chicago/UNITED STATES OF AMERICA, 4Pathology, The University Of was to investigate the expression and impact of FGFs and FGFRs in MPM Chicago, Chicago/IL/UNITED STATES OF AMERICA and to evaluate their potential suitability as new therapeutic targets.

Background: Prominent clinical activity of the PI3K/mTOR inhibitor GDC- Methods: Expression of all known FGF and FGFR genes was assessed by 0980 has been observed in patients with malignant mesothelioma (iMig qRT-PCR and confirmed by expression array analysis in MPM cell lines and abstract: 388). We investigated candidate mechanisms of PI3K inhibitor normal mesothelial cells. Selected FGFs/FGFRs were also evaluated on sensitivity in a panel of mesothelioma cell lines and tumor tissues. human tissue samples by immunohistochemistry. FGFR-specific tyrosine kinase inhibitors and an adenovirus construct expressing dominant- Methods: We screened an initial panel of thirteen epithelial, pleural negative FGFR1 were used to block FGF signal transduction in MPM mesothelioma tissue samples and eight epithelial mesothelioma cell lines cell models. The impact of FGFR inhibition as well as stimulation with for PIK3CA/PIK3R1 mutations (Sanger Sequencing), copy number alterations rhFGF2 on MPM cell proliferation, survival, apoptosis, cell cycle as well in 147 genes commonly altered in cancer (Nanostring nCounter). We as migration and invasion was evaluated by MTT, clonogenic, spheroid subsequently validated protein expression by immunoblotting (including formation, platypus, and transwell assays, video microscopy, and flow PTEN, INPP4B, NF2/Merlin, LKB1, NHERF1). In a second larger cohort of cytometry. The effect on downstream signal transduction was assessed by 20 mesothelioma cell lines, sensitivity to GDC-0980, and the presence of immunoblotting with phosphorylation site-specific antibodies. In addition, INPP4B, NF2, PIK3CA hotspot mutations, and other PI3K pathway related potentially additive or synergistic antineoplastic activity of FGFR1 inhibitors proteins were evaluated. We determined apoptosis by Annexin V/PI staining in combination with clinically applied chemotherapeutics, radiotherapy in a subset of cell lines. Cell line morphology in response to GDC-0980 was and other targeted drugs against MPM were investigated. An orthotropic examined by light microscopy. mouse model has been established for the evaluation of in vivo growth of MPM cells transduced with adenoviral constructs. Results: No canonical mutations in the PIK3CA/PIK3R1 genes were identified in any tumor sample or cell line. Several SNPs of unknown Results: Expression analysis revealed high expression of FGFR1, FGF2 significance were identified. Copy number analysis did not show loss and FGF18 in MPM cell lines as well as in tumor tissues. Stimulation with of either PTEN or NF2, however, copy number loss of tumor suppressor exogenous FGF2 led to a remarkable increase in cell migration and invasion INPP4B was seen in 6/13 (46%) tumor samples. Furthermore, this analysis accompanied by dramatic changes in cell morphology indicating EMT. In showed that copy number gain in commonly amplified oncogenes is very contrast, inhibition of FGFR1 by specific small molecule kinase inhibitors

iMig2012.org • Abstract Book 43 led to significantly decreased proliferation, survival, migration, invasion and spheroid formation in the majority of cell lines tested. Interestingly cell lines with intrinsic or acquired cisplatin resistance were more sensitive to FGFR inhibition. Adenoviral expression of dominant-negative FGFR1 further confirmed these results. FGFR1 inhibition further led to a G1 cell cycle arrest correlating with drug sensitivity. Additionally, when FGFR inhibition was combined with chemo-and radiotherapy, additive and synergistic effects were observed. Currently, an in vivo experiment with MPM cells expressing dominant-negative FGFR1 is performed. The results of these experiments will be presented at the conference.

Conclusion: Taken together, these data suggest that FGFR signals contribute to proliferation, survival, migration, invasion and chemo- and radiotherapy resistance of MPM cells and their inhibition should be further evaluated as a potential new treatment strategy in MPM. september 2012 13,

Disclosure: No significant relationships.

SESSION IVA GENE REGULATION AND MESOTHELIOMA PATHOGENESIS 3 September 13, 2012 14:20-16:00

IVA.7: CRITICAL PATHWAYS IN MALIGNANT MESOTHELIOMA

Marie-Claude Jaurand1, Didier Jean2 1Umr-S 674, Inserm, Paris/FRANCE, 2Inserm Umr-S 674, Inserm, Paris/ FRANCE

Tumor growth is controlled by numerous pathways, which are regulated by the activity of both intrinsic effectors and extrinsic factors. Present researches aim to identify the deregulated signaling pathways in malignant cells, with the goal to select molecules or mechanisms that could kill tumor cells or abolish tumor growth. In malignant mesothelioma (MM), the identification of gene mutations, epigenetic alterations and study of gene expression profiles has defined abnormalities and changes in cell homeostasis. Emphasis has been put on the deregulation of several pathways that can account for mesothelial cell neoplastic transformation. The most exciting alteration concerns the Hippo pathway, as about fifty percent of MM show a mutation of the NF2 gene. A number of studies have emphasized the role of receptor tyrosine kinase driven signaling, involving most of the growth factor receptors (VEGFR, IGFR, EGFR, cMET). The activation of MAPK (mitogen-activated protein kinase), phosphatidylinositol-3-kinase (PI3K-AKT), and the Wnt signaling pathway international mesothelioma interest group has been also reported. From our knowledge, these pathways are interconnected, and it is a challenge to choose the most critical pathways to target for a therapeutical efficiency. An approach can be accomplished by using one or more agents inhibiting a specific pathway. This strategy may also attenuate other signaling pathways. It is also possible to use agents to inhibit two or more signaling pathways. Several studies are in progress to determine the potency of these approaches. In a genetic and transcriptomic study of sixty MM cell lines, we identified several groups of MM showing distinct expression of genes involved in the epithelial- mesenchymal transition (D. Jean et al this meeting). Sub-groups also differ by the expression in effectors belonging to metabolic or growth factor pathways, showing the heterogeneity of MM. MM cells show alteration of several regulatory pathways regulating cell proliferation, apoptosis and survival, and adhesion and migration. To select the most appropriate anti-tumor agents, it is therefore important to determine the relationships between these pathways. Both the number of effectors altered and the extent of the connections between different pathways may be of importance. In any case, it is needed to identify biomarkers in tumor cells, and in microenvironment, indicative of the functional status of MM cells and predictive of the response to anti-tumor agents. These agents should be as specific as possible of the different subgroups of MM.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 44 Session IVB Novel Therapeutics: Clinical Trials September 13, 2012 14:20-16:00

SESSION IVB NOVEL THERAPEUTICS: CLINICAL TRIALS David Kwiatkowski8, Jennifer O. Lauchle9, Howard Burris3, Andrew September 13, 2012 14:20-16:00 Wagner6, Johan De Bono2 1

Section Of Hematology/Oncology, University Of Chicago, Chicago/IL/ september 2012 13, IVB.1: A PHASE II STUDY WITH THE ANTI-CTLA-4 MAB UNITED STATES OF AMERICA, 2The Institute Of Cancer Research And Royal TREMELIMUMAB IN CHEMOTHERAPY-RESISTANT ADVANCED Marsden Hospital, Sutton/UNITED KINGDOM, 3Sarah Cannon Research MALIGNANT MESOTHELIOMA Institute, Nashville/TN/UNITED STATES OF AMERICA,4Department Of Medicine, Thoracic Oncology Service, Memorial Sloan-Kettering Luana Calabro1, Aldo Morra2, Ramy Ibrahim3, Alessandra Di Pietro3, Cancer Center, New York/NY/UNITED STATES OF AMERICA, 5Radiology, Diana Giannarelli4, Luciano Mutti5, Michele Maio1 Columbia University Medical Center, New York/NY/UNITED STATES OF 1Medical Oncology And Immunotherapy - University Hospital Of Siena, AMERICA, 6Dana Farber Cancer Institute, Boston/MA/UNITED STATES OF Siena/ITALY, 2Dept Of Radiology,, Euganea Medica Diagnostic Center, AMERICA, 7Radiology Department, Royal Marsden Hospital, Sutton/UNITED Padua, Italy, Padua/ITALY, 3Clinical Development Oncology, Medimmune, KINGDOM, 8Brigham And Women’s Hospital, Boston/MA/UNITED STATES Gaithersburg/UNITED STATES OF AMERICA, 4Istituto Tumori-Regina Elena, OF AMERICA, 9Exploratory Clinical Development, Genentech, South San Rome/ITALY, 5Dept. Of Medicine, Vercelli Hospital, Vercelli/ITALY Francisco/CA/UNITED STATES OF AMERICA

Background: Anti-CTLA-4 monoclonal antibodies (mAb) are showing Background: The PI3K-AKT-mTOR signaling pathway is dysregulated in significant activity in different tumor types; however, no data are available many cancers, including MPM. GDC-0980 is a potent, selective, oral in MM patients (pts). We report results of a phase II, single Institution, inhibitor of class I PI3K and mTOR kinase with in vitro IC50 of 4.8 nM for study investigating safety, clinical and immunologic efficacy of the fully- p110a/p85a and apparent Ki of 17.3 nM for human mTOR. human anti-CTLA-4 mAb tremelimumab as second-line treatment for advanced MM pts. Methods: A phase I dose-escalation study using a 3+3 design was initiated at centers in the United States and United Kingdom in order Methods: Second-line advanced mesothelioma pts, who relapsed after to determine the safety and tolerability of GDC-0980 in patients with a prior platinum-based regimen, were enrolled in the study and received solid tumors. Preliminary efficacy was assessed, and pharmacokinetic, tremelimumab at 15 mg/kg i.v. on day (d) 1 and 90 for 4 cycles or until pharmacodynamic, and biomarker studies were performed. Based on progressive disease (PD), or unacceptable toxicity. Primary endpoint tumor shrinkage observed in MPM in the dose-escalation portion of was objective response (OR); secondary endpoints were safety, disease the study, an expansion cohort of approximately 20 MPM patients was control rate (DCR), overall survival (OS), and immunologic activity. Tumor initiated at 30-mg GDC-0980 daily. CT scans were centrally reviewed assessment per modified RECIST Criteria was performed at screening and retrospectively by a radiologist with MPM expertise. Archival tumor at d 80 of each cycle. Adverse events (AE) were collected according to the tissue was evaluated for PIK3CA mutation by allele specific PCR or Sanger Common Terminology Criteria v3.0. Peripheral blood mononuclear cells sequencing; PTEN expression was assessed by immunohistochemistry.

were collected at baseline, d 14, 30, 60, and 90 of each cycle and were international mesothelioma interest group analyzed by flow cytometry for an extensive panel of immune phenotypic Results: Six MPM patients were enrolled in the dose-escalation portion and T-cell activation markers. of the study. Anti-tumor activity was observed in 5 MPM patients at GDC-0980 doses of 8 to 50 mg, with tumor shrinkage of 5 to 36% per Results and Conclusions: From May 2009 to January 2012, 29 advanced modified RECIST. Two patients achieved a partial response, including MM pts were enrolled and received at least 1 dose of tremelimumab one with a tumor containing a PIK3CA mutation (exon 2 R88Q) treated at (median 2; range 1-7). Preliminary results show that Tremelimumab is 8 mg. Three patients experienced drug-related adverse events that led active in MM pts, and can induce durable stabilization of disease in a to treatment discontinuation before completing cycle 2: 1 patient had significant proportion of pts, warranting further investigation. The AEs grade 3 hyperglycemia at the 70 mg dose; 2 patients at the 40 mg dose observed in this study are consistent with Tremelimumab safety profile in developed grade 3 or greater pneumonitis/pneumonia (including one fatal other indications. Treatment associates with major changes in activated case). Efficacy and safety data are currently available for 7 of 20 patients and memory T cell subpopulations.. In light of these results, the Study treated with 30 mg of GDC-0980 in the MPM expansion cohort. Reported MESOT-TREM-2012 is currently ongoing to explore a different schedule of drug-related Grade 1-2 adverse events were similar to the general study treatment with tremelimumab in advanced refractory MM pts. population; one Grade 3 event of rash was reported. Anti-tumor activity has been observed: maximum tumor change from baseline ranges from Disclosure: No significant relationships. +17% to -21%; 2 of the 7 MPM patients remain on study for >6 months, and 3 patients remain on study with 2-3 month follow-up.PIK3CA mutations were uncommon; loss of PTEN expression was not observed.

SESSION IVB NOVEL THERAPEUTICS: CLINICAL TRIALS Conclusion: GDC-0980 at 30 mg daily is generally well-tolerated in September 13, 2012 14:20-16:00 MPM patients. Anti-tumor activity, evidenced by tumor regression and prolonged disease control, has been observed. PIK3CA mutations were IVB.2: EVALUATION OF TOLERABILITY AND ANTI-TUMOR uncommon; PTEN null tumors were not observed. Updated data on clinical ACTIVITY OF GDC-0980, AN ORAL PI3K/MTOR INHIBITOR, outcomes and biomarker correlates in the remaining patients in the ADMINISTERED DAILY IN PATIENTS WITH ADVANCED MALIGNANT expansion cohort will be presented. PLEURAL MESOTHELIOMA (MPM) Disclosure: Consultant/advisor, Genentech, compensated: HL Kindler, J Hedy L. Kindler1, Saorise Dolly2, Johanna Bendell3, Lee M. Krug4, deBono, LM Krug, A Wagner Employment, Genentech: JO Lauchle Lawrence Schwartz5, Michael Rabin6, Nina Tunariu7, Tanguy Y. Seiwert1, Marjorie G. Zauderer4, Ann M. Young2, Jennifer Shouldis1, J P. Marcoux6,

iMig2012.org • Abstract Book 45 SESSION IVB NOVEL THERAPEUTICS: CLINICAL TRIALS Cancer Centre, Adelaide/SA/AUSTRALIA September 13, 2012 14:20-16:00 Background: BNC105P is a tubulin polymerization inhibitor that acts as IVB.3: PHASE II TRIAL OF ANTI-TRANSFORMING GROWTH a Vascular Disruption Agent (VDA), has direct cytotoxic effects, and had FACTOR-BETA (TGFß) MONOCLONAL ANTIBODY GC1008 IN preclinical and phase I activity in malignant pleural mesothelioma (MPM). RELAPSED MALIGNANT PLEURAL MESOTHELIOMA (MPM) This aim of this study was to determine activity, safety, and potential biomarkers of BNC105P as second line therapy after pemetrexed and a James Stevenson1, Hedy Kindler2, Daniel Schwed3, platin in MPM. Anjana Ranganathan3, Mona Jacobs-Small3, Jennifer Shouldis2, Steven M. Albelda3 Methods: Eligible patients had progressive MPM, prior pemetrexed and 1Taussig Cancer Institute, Cleveland Clinic Foundation, Cleveland/OH/ platinum, measurable disease by modified RECIST, ECOG PS 0-1, and UNITED STATES OF AMERICA, 2University Of Chicago, /UNITED STATES adequate organ and cardiovascular function. Important exclusions included OF AMERICA, 3Medicine, University Of Pennsylvania/UNITED STATES OF recent thromboembolic, cardiovascular or cerebrovascular disease, or AMERICA therapeutic anticoagulation. Patients received BNC105P (16 mg/m2 I.V) Day 1 + 8 q21d until progression or prohibitive toxicity. The primary endpoint september 2012 13, Background: TGFβ is a pleiotropic cytokine overexpressed by MPM. Based was centrally reviewed objective tumour response rate (RR = CR + PR); on preclinical data documenting a key role for TGFβ in promoting growth the Simon 2-stage design assumed a RR of interest of 20% and a RR of no and progression of MPM, we are conducting a Phase II trial of GC1008 in interest of 5%, with α = β = 0.05. Continuation past first stage accrual patients (pts) with progressive MPM. required >1 objective response in 24 patients. Serum mesothelin was measured on day 1 of each cycle. A panel of 62 potential biomarkers of Methods: Pts with progressive MPM by modified RECIST criteria and PS response were measured at baseline, 3 hours after the first dose of study 0-1 with 1-2 prior systemic therapies (at least 1 pemetrexed-based) are drug, and on day 8 cycle 1. eligible. Treatment plan: GC1008 3mg/kg IV over 90 minutes every 21 days. Responses are assessed by modified RECIST every 6 weeks. The primary Results: 30 subjects were accrued over 10 months (90% male; median endpoint is progression-free survival (PFS) rate at 3 months; secondary age 65 (range 41-83); 77% ECOG PS 1; histology epithelioid (67%), objectives include safety with GC1008, response rate by modified RECIST, biphasic (10%), sarcomatoid (7%), other/unspecified (17%)). All patients time to progression (TTP), and overall survival (OS). received at least one dose of study drug; patients received a median of 2 cycles and median dose intensity was 100%. No significant haematologic, Results: The modified Gehan stage 1 stopping criterion of 1/13 pts with 3 biochemical, peripheral neurotoxic or cardiac adverse events (AEs) month PFS has been exceeded. To date, 13 pts (10 PS 0; 3 PS 1) with MPM including hypertension were observed. Grade 3 or 4 AEs occurred in 10 (median age 69; 2F, 11M; 11 epithelial, 1 sarcomatoid, 1 biphasic) enrolled. pts (33%). There were 2 deaths on study: 1 due to stroke, the other due Treatment-related toxicities include G1/2 fatigue (3 pts), nausea (1 pt) and to pneumonia and respiratory failure. We observed 1 partial response (PR) xerosis (1 pt). Other adverse events possibly related to GC1008 were rapid (3%) and 13 pts with stable disease as their best response (43%). Median disease progression in 1 pt after 2 cycles, and G2 skin keratoacanthoma progression free survival was 1.5 months (95% CI 1.4-2.4); median overall in 1 pt after 5 cycles. Three pts met the primary objective of 3 month PFS survival was 8.7 months (95% CI 3.8-NR). One patient, who acheived a at 4.1, 4.2 and 9 months each. Stable disease (SD) was seen in 3 pts (23). PR, had a 25% decrease in mesothelin levels after the first treatment cycle. Median TTP is 1.4 months (95% CI 1.2-∞); median OS is 13 months (95% Significant elevations were observed in MIP-1beta, Ferritin, IL-8, IL-10, CI 6-∞). Increased serum mesothelin levels have closely tracked disease TNFR2, and IL-16 following the first dose of study drug. Lung function and progression. Serum from 6/13 pts showed new antibodies against MPM quality of life data will also be presented. tumor lysates as measured by immunoblotting. Two of 3 pts with SD had anti-tumor antibody responses. Mean baseline plasma level of TGFβ was Conclusion: BNC105P was safe and tolerable but its single agent response 2447 pg/ml but did not correlate with baseline plasma TGFβ or T T P. rate failed to meet our pre-specified criterion. Mesothelin response was concordant with radiological response. Post-dose biomarker alterations Conclusions: GC1008 was well tolerated in pretreated MPM patients. SD demonstrated a pharmacodynamic effect consistent with endothelial stress international mesothelioma interest group occurred in 3 pts, all with prior disease progression. Evidence for humoral but insufficient objective radiological responses were seen to correlate this anti-tumor immunity was seen in nearly half of enrollees and in 2 of 3 pts with outcomes. with SD. OS compares favorably to prior single-agent studies in pretreated MPM. Disclosure: Bibby, Doolin, Kremmidiotis, Lavranos: Employment by Bionomics Pty Ltd, developers of BNC105P. Note that these authors have Disclosure: No significant relationships. had the opportunity to review results and contribute to the abstract, however did not have the ability to veto any abstract content

SESSION IVB NOVEL THERAPEUTICS: CLINICAL TRIALS September 13, 2012 14:20-16:00 SESSION IVB NOVEL THERAPEUTICS: CLINICAL TRIALS September 13, 2012 14:20-16:00 IVB.4: PHASE II TRIAL OF BNC105P AS 2ND LINE CHEMOTHERAPY FOR ADVANCED MALIGNANT PLEURAL MESOTHELIOMA (MPM). IVB.5: ARGININE DEPRIVATION WITH PEGYLATED ARGININE (AUSTRALASIAN LUNG CANCER TRIALS GROUP AND NHMRC DEIMINASE (ADI-PEG20) IN PATIENTS WITH MALIGNANT CLINICAL TRIALS CENTRE COLLABORATION) PLEURAL MESOTHELIOMA (MPM): PRELIMINARY RESULTS OF A MULTICENTER RANDOMIZED PHASE II TRIAL Anna K. Nowak1, Chris Brown2, Michael J. Millward1, Brett Hughes3, David Bibby4, Gabriel Kremmidiotis4, Elizabeth Doolin4, Paul Mitchell5, Peter Szlosarek1, Jeremy P. Steele1, Michael Sheaff2, Norbert Avril3, Nick Pavlakis6, Xanthi Coskinas2, Lucille Sebastian2, Michelle Cummins2, Teresa Szyszko3, Stephen Ellis4, Luke Nolan5, Paul Taylor6, Michael J. Jenette Creaney1, Michael Boyer7, Francis Parnis8, Tina Lavranos4, Lind7, David Gilligan8, James Spicer9, Melissa Phillips1, Fiona Luong10, Martin Stockler2 Nicholas Lemoine11, John Bomalaski12, Robin Rudd1, Dean Fennell13, Allan 1School Of Medicine And Pharmacology, University Of Western Australia, Hackshaw14 Perth/AUSTRALIA, 2Nhmrc Clinical Trials Centre, University Of Sydney, 1Department Of Medical Oncology, St Bartholomew’s Hospital, London/ Sydney/NSW/AUSTRALIA, 3Medical Oncology, The Prince Charles UNITED KINGDOM, 2Department Of Histopathology, St Bartholomew’s Hospital, Brisbane/QLD/AUSTRALIA, 4Bionomics Pty Ltd, Adelaide/SA/ Hospital, London/UNITED KINGDOM, 3Department Of Nuclear Medicine, AUSTRALIA, 5The Austin Hospital, Melbourne/VIC/AUSTRALIA, 6Medical St Bartholomew’s Hospital, London/UNITED KINGDOM,4Department Oncology, Royal North Shore Hospital, /NSW/AUSTRALIA, 7Medical Of Radiology, St Bartholomew’s Hospital, London/UNITED Oncology, Royal Prince Alfred Hospital, Sydney/NSW/AUSTRALIA, 8Adelaide KINGDOM, 5Medical Oncology, Southampton General Hospital/UNITED

iMig2012.org • Abstract Book 46 KINGDOM, 6Wythenshawe Hospital/UNITED KINGDOM, 7Postgraduate specific anti-tumor responses with minimal “on target, off tumor” effects. Medical Institute, University Of Hull/UNITED KINGDOM, 8Addenbrooke’s We have generated a mesothelin chimeric antigen receptor (CAR) which Hospital/UNITED KINGDOM, 9Department Of Oncology And Hematology, modifies T cells to target mesothelin with antibody specificity coupled Guy’s Hospital/UNITED KINGDOM, 10Molecular Oncology, Barts with T cell signaling and cytolytic effect without MHC restriction. In murine Cancer Institute, Queen Mary University Of London, London/UNITED MPM models, lentiviral and mRNA transduced mesothelin CAR T cells KINGDOM, 11Cancer Services, St Bartholomew’s Hospital, London/ have dramatic and persistent antitumor effects in large tumors with no UNITED KINGDOM, 12Polaris Group, San Diego/UNITED STATES OF associated serosal or other toxicity. Based on these data, we designed a AMERICA, 13University Of Leicester/UNITED KINGDOM, 14Ucl Cancer human phase I dose escalation trial to test the safety and manufacturing Research Uk Clinical Trials Center/UNITED KINGDOM feasibility of mRNA transduced mesothelin CAR T cells. The mRNA approach was undertaken as a safety measure as vector genomic integration cannot Background: Preclinically arginine deprivation has shown activity as a occur and if serious or dose-limiting toxicity were to develop, mesothelin novel antimetabolic strategy for MPM lacking the rate-limiting enzyme CAR expression and associated toxicity should be transient. for arginine biosynthesis argininosuccinate synthetase (ASS1). We have initiated a multicenter randomized phase II Cancer Research UK-funded Methods: Eligible patients undergo apharesis and lymphocytes are clinical trial to assess the safety and efficacy of the arginine-lowering agent transfered to the Cell and Vaccine Production Facility to undergo september 2012 13, ADI-PEG20 (Polaris Group, San Diego, US) and best supportive care (BSC) electroporation with the mesothelin CAR construct and expansion with versus BSC alone in patients with ASS1-deficient MPM. CD3/28 beads. After electroporation and expansion, cells are frozen in batches for infusion. As a further safety measure in addition to the mRNA Methods: Eligibility criteria for the sixty-six patients include: non- approach, a two cohort design was undertaken. Cohort one receives a resectable, ASS1-deficient MPM by immunohistochemistry; good single infusion of 1e8 mesothelin CAR T cells followed one week later by performance status (0 or 1); measurable disease by modified RECIST 1e9 mesothelin CAR T cells. Once safety is demonstrated in a typical 3-3 (Response Evaluation Criteria in Solid Tumors); and able to give written design, cohort 2 patients will receive one cycle of 1e8 CAR T cells (one cycle consent. Stratification factors include: gender, histology (sarcomatoid defined by Mon, Wed, Fri T cell infusion), followed 2 weeks later by one versus non-sarcomatoid), prior treatment (chemonaive or previous cycle of 1e9 CAR T cells. Serologic samples are collected to look for CAR platinum combination therapy), and recruitment center. The primary expression over time, to assess immunologic alterations, and to assess for endpoint is progression free survival (PFS) as assessed using modified human anti-mouse humoral responses. Repeat imaging to assess response RECIST. Secondary endpoints are tumor response rate, overall survival occurs 35 days post CAR T cell infusion. and toxicity. Tertiary endpoints include measurement of plasma arginine, citrulline and ADI-PEG20 antibody levels, the methylation status of Results: Three patients have completed cohort 1 without serious adverse the ASS1 gene in primary tumoral samples, and early metabolic response to event or dose limiting toxicity. Two patients had no T cell infusion related ADI-PEG20 using [18F]Fluorodeoxyglucose Positron Emission Tomography side effects, one patient had mild cytokine release symptoms. Based on (FDG-PET). Sample size was calculated to detect a hazard ratio of 0.60 FDA approval, one patient proceeded into cohort 2 and demonstrated (80% power, 15% one-sided significance level). similar mild cytokine release symptoms as he had in cohort 1. Serum mesothlin CAR expression peaks after T cell infusion and then progressively Results: To date up to 50% of patients have been screened ASS1-deficient, decline over several days. In addition, atlerations in the pro-inflammatory and over half of patients have been randomized onto the trial which is cytokine milieu were seen and one patient developed anti-mouse projected to complete accrual by the end of 2012. ADI-PEG20 toxicity in antibodies against a region of the CAR construct. Radiographic progression patients with MPM has been consistent with previous trials of ADI-PEG20 was seen in all 3 patients at day 30 by traditional radiographic measures. in melanoma and liver cancer, commonly skin injection site reactions (grade 1-2), infrequent episodes of neutropenia (range: grade 1-4) and Conclusion: We have been able to demonstrate preliminary safety and anaphylactoid reactions (2 patients with grade 3 episodes). Preliminary manufacturing ability of mesothelin CAR T cells in patients with MPM. Upon metabolic response data indicate that FDG-PET imaging may be a useful completion of this phase I study and safety confirmation, we plan to move tool to assess early response to ADI-PEG20 in patients with loss of tumoral forward with lymphodepletion and/or lentiviral transduced T cells in order expression of ASS1. The trial has passed the first interim safety analysis and to enhance T cell persistence and anti-tumor effect. international mesothelioma interest group an update will be provided on current recruitment, safety and response as assessed by ASS1 promoter methylation status and FDG-PET. Disclosure: No significant relationships.

Conclusion: In summary, ADI-PEG20 is well tolerated and shows evidence of metabolic response in patients selected for ASS1-deficient MPM. Arginine deprivation may have a role in the future management of MPM SESSION IVB NOVEL THERAPEUTICS: CLINICAL TRIALS either alone or in combination with rationally selected therapies. September 13, 2012 14:20-16:00

Disclosure: No significant relationships. IVB.7: MEASLES VIROTHERAPY OF MESOTHELIOMA: UPDATE ON PHASE I CLINICAL TRIAL AND THE POTENTIAL OF ENDOTHELIAL CELL CARRIERS TO EVADE ANTIVIRAL IMMUNE RESPONSE.

SESSION IVB NOVEL THERAPEUTICS: CLINICAL TRIALS Manish R. Patel1, Blake A. Jacobson2, Evanthia Galanis3, Tobias Peikert4, September 13, 2012 14:20-16:00 Robert P. Hebbel1, Stephen J. Russell5, Robert A. Kratzke2 1Medicine, Division Of Hematology, Oncology, And Transplantation, IVB.6: A PHASE I TRIAL OF MESOTHELIN CHIMERIC ANTIGEN University Of Minnesota Medical School, Minneapolis/MN/UNITED RECEPTOR T CELLS IN PATIENTS WITH ADVANCED PLEURAL STATES OF AMERICA, 2Medicine, Div Of Hematology, Oncology, And MESOTHELIOMA. Transplantation, University Of Minnesota, Minneapolis/MN/UNITED STATES OF AMERICA, 3Oncology, Mayo Clinic, Rochester/MN/UNITED STATES OF Andrew R. Haas, Edmund Moon, Adri Recio, Daniel Sterman, Michael AMERICA, 4Pulmonary And Critical Care Medicine, Mayo Clinic, Rochester/ Kalos, Carl June, Steven M. Albelda MN/UNITED STATES OF AMERICA, 5Hematology, Mayo Clinic, Rochester/ University Of Pennsylvania Medical Center, Philadelphia/PA/UNITED STATES MN/UNITED STATES OF AMERICA OF AMERICA Background: Edmonston-strain measles virus (MV) is a promising Background: Malignant pleural mesothelioma (MPM), particularly oncolytic virus. Preliminary data shows that MV has oncolytic activity on epithelioid subtype, demonstrates high cell surface mesothelin expression mesothelioma cells in vitro and on human xenografted tumors. Therefore, relative to normal tissues where expression is limited to the pleural, a phase I clinical trial of intrapleural administration of MV was initiated for peritoneal, and pericardial serosal surfaces. This characteristic makes patients with mesothelioma. In order to better understand the mechanisms mesothelin an ideal target for immunotherapeutic approaches to maximize of resistance and sensitivity, experiments were planned to study the

iMig2012.org • Abstract Book 47 immune response to measles virus as well as tumor characteristics that might account for antitumor activity. One of the major limitations to virotherapy is the generation of neutralizing antibodies. Since >80% of the population is vaccinated against MV, this could potentially be a limitation of MV therapy for mesothelioma. Using ex vivo, infected cell carriers can overcome this limitation by hiding the virus within intact cells allowing for trafficking of virus toward the tumor and oncolytic activity. Blood outgrowth endothelial cells (BOEC) are easily obtained from a blood sample, are readily cultured, and have tumor homing characteristics. Therefore, cell carrier mediated measles virotherapy is tested as a potential novel mechanism for delivery of MV to mesothelioma.

Methods: For the phase I clinical trial, all patients with malignant mesothelioma able to have a pleural catheter placed are eligible regardless of line of therapy. Patients having prior pleurodesis or a current indwelling september 2012 13, pleural catheter for greater than 6 weeks are excluded. MV-expressing sodium-iodide symporter (MV-NIS) will injected into the pleural space in a dose-escalation schema. Radioactive iodine uptake will be a measure of viral replication as well as viral titers from blood and pleural fluid. Correlative studies to assess immune response and expression of potential biomarkers will be performed. BOECs were cultured from human donor blood, cultured, and characterized using immunofluorescence for endothelial markers. 150,000 BOECs were infected, ex vivo with MV expressing GFP or CEA (MV-GFP or MV-CEA) and monitored for cell survival at various times. MV-infected BOECs were also co-cultured with human mesothelioma cell lines in the presence or absence of MV immune serum. Viability of mesothelioma cells after 72 hours of co-culture were determined by trypan blue exclusion. Animal experiments are planned to test the efficacy of the cell carrier approach using a peritoneal model of xenografted human mesothelioma. Bioluminescence of xenografted mesothelioma cells expressing firefly luciferase will be used to gauge antitumor response. Results will be compared to administration of naked virus and untreated controls.

Results: For the phase I study, one patient has been enrolled in the study. At this time, the patient has completed one treatment and it is too early to assess treatment response or to complete correlative studies. BOECs are readily infected by MV and most cells survive infection up to 72 hours with an MOI of 1. Cytopathic effect can be visualized by microscopy by 48 hours. Co-culture of BOECs with mesothelioma cells results in complete oncolysis of mesothelioma cells.

Conclusion: Phase I study of MV therapy is ongoing. Too few patients have been treated to draw conclusions. Further investigation of BOECs as a international mesothelioma interest group delivery method for MV to mesothelioma is warranted.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 48 Session IVC Pathology September 13, 2012 14:20-16:00

SESSION IVC pATHOLOGY in male p=0.006, older, while rich myxoid type was most often observed September 13, 2012 14:20-16:00 in female ( p=0.065) and younger age(p=0.0015). There were no significant

difference between asbestos and non asbestos exposed patients. september 2012 13, IVC.2: PROGNOSTIC SIGNIFICANCE OF HISTOLOGICAL SUBTYPES IN PLEURAL MALIGNANT MESOTHELIOMA ; A Conclusion: Our study support classification by histological type CLINICOPATHOLOGICAL REVIEW AND A PROPOSAL FOR and subtype for a better stratification of patient survival. Molecular UPDATING HISTOLOGICAL MESOTHELIOMA CLASSIFICATION. classification in the future will evaluate the strenght of such indicators.

Francoise Galateau Sallé1, Nolwenn Le Stang2, Maria Paciencia3, Marie Disclosure: No significant relationships. Karanian3, Alan Borsczuck4, Kouki Inai5, Allen Gibbs6, Junya Fukuoka7, Kenzo Hiroshima8, Marleen Praet9, Jean Michel Vignaud1 1Chu Caen, Mesopath Im@Ec, Caen Cedex/FRANCE, 2Pathology Department, Mesonat Registry, Caen Cedex 9/FRANCE, 3Pathology SESSION IVC pATHOLOGY Departement - University Hospital, U 1086 Inserm “Cancers & September 13, 2012 14:20-16:00 Préventions”, Caen Cedex 9/FRANCE, 4Anatomic Pathology, Columbia University/UNITED STATES OF AMERICA, 5Pathology, Nstitute Of Biomedical IVC.3: DIAGNOSTIC UTILITY OF P16 FISH IN DISTINCTION & Health Sciences Hiroshima University, Hiroshima/JAPAN, 6Llandough BETWEEN SARCOMATOID MESOTHELIOMA AND FIBROUS University, Cardiff/UNITED KINGDOM, 7Mesopath Im@Ec, Caen Cedex/ PLEURITIS FRANCE, 8Chiba University, Chiba/JAPAN, 9Gent University, Gent/BELGIUM Kenzo Hiroshima1, Di Wu1, Shinji Matsumoto2, Kazuki Nabeshima2, Background: Age, epithelioid type, T and N staging are conventional Toshikazu Yusa3, Daisuke Ozaki3, Michio Fujino4, Yukio Nakatani5, Yuji prognostic factors in diffuse pleural malignant mesothelioma [DPMM] Tada5, Hideaki Shimada6, Masatoshi Tagawa7 useful for the management of the patients. Recent results have focused 1Department Of Pathology, Tokyo Women’s Medical University, Yachiyo/ some interest on whether histological subtyping among epithelioid and JAPAN, 2Pathology, Fukuoka University, Fukuoka/JAPAN, 3Chiba Rosai sarcomatoid DPMM is a prognostic factor and allow a better stratification Hospital, Ichihara/JAPAN, 4Chiba East Hospital, Chiba/JAPAN, 5Chiba of patients for survival. Our aim was to assess wether overall survival of University, Chiba/JAPAN, 6Department Of Surgery, Toho University School patients depends on histological subtyping. Of Medicine, Tokyo/JAPAN, 7Division Of Pathology And Cell Therapy, Chiba Cancer Center Research Institite, Chiba/JAPAN Methods: We undertook a retrospective study and reveiwed H&E slides of mesothelioma cases from the MESOPATH domestic referal center files Background: Fibrous pleuritis is sometimes difficult to distinguish from during 1995-2010. They were thoracoscopy biopsy in 40% (n=206) and sarcomatoid mesothelioma, especially when the amount of tumor in the surgical specimens in 60% (n=323). We classified the tumors according biopsy is small. There are some immunohistochemical markers which are international mesothelioma interest group to the WHO 2004 classification and we subclassified the epithelioid useful for the differential diagnosis between epithelioid mesothelioma type in papillary, trabecular, acinar, solid, micropapillary. Additionnally and reactive mesothelial hyperplasia. However, immunohistochemistry we evaluate the lymphohistiocytoid, the pleomorphic and an additional is inconclusive for the differential diagnosis between sarcomatoid epithelioid variant with rich mxoid stroma. Log rank test and cox models mesothelioma and fibrous pleuritis. Tumor invasion of the stroma is the were used to analyse the relation between histological type and subtype most reliable criteria for the diagnosis of sarcomatoid mesothelioma; variables, clinical features, asbestos exposure and survival. however, it cannot be assessed in small biopsies. It is reported that most of mesothelioma patients have deletion of p16 and methylated p16 promoters. Results: From a population study of 526 patients, there were 381 male In this study, we studied the diagnostic utility of p16 FISH and methylation (72%) and 145 female (28%) with an average age of 70 years old [28;96]. status of p16 for the diagnosis of malignant pleural mesothelioma (MPM). Asbestos exposure was observed in 71 % of cases). The histological types and subtypes was papillary 5% (n=25), solid 16%(n=83), trabecular 21% Methods: A retrospective analysis of 48 surgical biopsies from patients (n=109), acinar 6% (n=30), micropapillary 7%(n=39), epithelioid type with MPM (17 epithelioid, 9 biphasic, 22 sarcomatoid) was performed. with rich myxoid stroma 6%(n=31), pleomorphic variant 12% ( n=65), Eighteen cases were treated with extrapleural pneumonectomy (EPP) Lymphohistiocytoid variant 7%( n=39) sarcomatoid (10%( n=53) and and 30 were treated with therapy other than EPP. The diagnosis desmoplastic type 10%(n=52) . We excluded from the study the biphasic of mesothelioma was rendered based on histology, results of type. Using age-adjusted Cox model, patients with desmoplastic type had immunohistochemistry, image studies, and clinical course. FISH for the worst prognosis with a 5 months median survival and a 2 years survival deletions of p16 was performed in 43 cases of mesothelioma and in 9 cases at 6% (p<0.0001) followed by the sarcomatoid and pleomorphic variants of fibrous pleuritis with benign asbestos pleural effusion. Methylation with a 5 months median survival but respectively a 2 years survival at 4% specific PCR were attempted in 35 cases. (p<0.0001) and 10% (p <0.0001). Lymphohistiocytoid type showed a 8 months median survival with 15% at 2 years survival (p=0.0003). Among Results: Homozygous and hemizygous deletion patterns were observed the epithelioid type micopapillary subtype presented the worst prognosis in mesotheliomas, but most of mesothelioma showed deletion patterns with 13 months median survival and 26 % at 2 years survival (p<0.07) in some of tumor cells. Homozygous deletion pattern was also observed followed by solid type 14 months. There was no significant statistical in some cells in fibrous pleurits due to artifactual cutting of nuclei. The difference of median survival for the trabecular and acinar types with mean percentage plus four standard deviations of cells with homozygous respectively a median survival at 16 and 18 months and 32% and 38% deletion pattern in fibrous pleurits was 15%. If a data distribution is at 2 years survival. The epithelioId type with rich myxoid stroma showed approximately normal, 99.994 percent of the data values are within four a 19 months median survival with 37% at 2 years survival (p<0.07) and standard deviations. We set the cut-off value as 15% for the diagnosis of finally papillary subtype showed the best prognosis with 21 months median MPM. The percentage of homozygous deletion pattern was higher than survival and 44% at 2 years survival. Noticeably, DMM was more present this value in 61.5% of the epithelioid mesotheliomas (8/13) and in all of the

iMig2012.org • Abstract Book 49 sarcomatoid mesotheliomas (22/22). Mesothelioma without homozygous Pathologically deletion showed tubulo-papillary pattern. Methylation of p16was observed confirmed MPM in 7 of 34 informative cases (21%). Mesotheliomas with homozygous deletion tend to be without methylation; however, two mesotheliomas + - TOTAL with homozygous deletion have been methylated. Mesotheliomas without Cytological + 283 2 285 deletion also tend to be without methylation. Furthermore, methylation diagnosis of p16 gene was observed in 43% of fibrous pleuritis (3/7). The presence of MPM of p16 homozygous deletion and p16 hypermethylation correlates with a shorter survival in patients with MPM. - 112 4860 4972 TOTAL 395 4862 5275 Conclusion: FISH analysis demonstrated homozygous deletion of the p16 in all of sarcomatoid mesothelioma cases (100%). Methylation of p16 was Sensitivity 72% Specificity 99% observed in 21% of MPMs. FISH analysis can identify both homozygous and hemizygous deletions. Our cut-off value for the diagnosis of mesothelioma Conclusion: The value of cytology in the diagnosis of pleural epithelioid is lower than previously reported. However, each institute should establish MM is unquestionable. Cytology provides a rapid, cheap and minimally september 2012 13, its own cut-off value. Different component in the same tumor can be invasive diagnostic modality, as well as enabling early diagnosis. Effusion simultaneously analyzed with p16 FISH. p16 FISH analysis can be a reliable cytology is rarely helpful in the diagnosis of sarcomatoid MM, and there are test for the distinction between sarcomatoid mesothelioma and fibrous some cases of epithelioid/biphasic MM which require biopsy to establish a pleuritis and for the diagnosis of sarcomatoid mesothelioma with unusual definitive diagnosis. histological finding and predict the prognosis of the patients. Disclosure: No significant relationships. Disclosure: No significant relationships.

SESSION IVC pATHOLOGY SESSION IVC pATHOLOGY September 13, 2012 14:20-16:00 September 13, 2012 14:20-16:00 IVC.5: PRE-OPERATIVE ANEMIA AS A PROGNOSTIC BIOMARKER IVC.4: A TWENTY YEAR AUDIT OF THE CYTOLOGICAL DIAGNOSIS FOR TIME TO RECURRENCE (TTR) AND OVERALL SURVIVAL (OS) OF PLEURAL MALIGNANT MESOTHELIOMA FOLLOWING EXTRA-PLEURAL PNEUMONECTOMY (EPP) AND PLEURECTOMY/DECORTICATION (PD) FOR MALIGNANT PLEURAL Amanda Segal1, Greg F. Sterrett1, Felicity A. Frost1, Anna K. Nowak2, MESOTHELIOMA (MPM) Arthur W. Musk3, Bruce W.S. Robinson4, Jenette Creaney2 1Pathwest, Nedlands/AUSTRALIA, 2School Of Medicine And Pharmacology, Elizabeth H. Baldini1, Ritu R. Gill2, Andrea S. Wolf3, Brian M. Goodman4, University Of Western Australia, Nedlands/AUSTRALIA, 3Respiratory Raphael Bueno4, Olivia Winfrey4, Hannah Eisen4, Juliann Barlow4, David Medicine, Sir Charles Gairdner Hospital, Nedlands/WA/AUSTRALIA, 4School J. Sugarbaker4, William G. Richards3 Of Medicine And Pharmacology, University Of Western Australia, Perth/ 1Radiation Oncology, Brigham And Women’s Hospital/Dana-Farber Cancer WA/AUSTRALIA Institute, Boston/MA/UNITED STATES OF AMERICA, 2Radiology, Brigham And Women’s Hospital, Boston/MA/UNITED STATES OF AMERICA, 3Division Background: Cytological examination of effusions provides an opportunity Of Thoracic Surgery, Brigham And Women’s Hospital And Harvard Medical to diagnose MM. However this is controversial and is not accepted School, Boston/MA/UNITED STATES OF AMERICA, 4Division Of Thoracic according to the current guidelines. In order to examine the role of Surgery, Brigham And Women’s Hospital, Boston/MA/UNITED STATES OF cytology in this diagnostic setting we conducted an audit of all pleural AMERICA cytology specimens received in our laboratory over a 20 year period. international mesothelioma interest group Background: Our recent research (Gill, AJR 2012) demonstrated that tumor Methods: All pleural specimens received from 1988-2007 were identified. associated anemia is an independent adverse prognostic factor for OS in Follow-up to establish final diagnosis was by review of pathology samples patients undergoing EPP for epithelial MPM. We further investigate this and data-linkage with Cancer and Mesothelioma Registries. association in relation to Common Terminology Criteria (CTC) grades of anemia, TTR, and whether this relationship also applies to non-epithelial Results: 9985 pleural samples were received (1740 biopsy, 8245 cytology) MPM and/or to patients undergoing PD. from 6198 individuals during the twenty year period. Approximately 30% of cytology samples were reported as either normal or non-diagnostic; 30% Methods: Retrospective review of 230 patients with mesothelioma who as inflammatory or as reactive hyperplasia; and 17% as metastatic to the underwent EPP or PD at Brigham and Women’s Hospital between 2001 and pleura or adenocarcinoma. Six percent of all pleural cytology samples over 2010 was performed. Demographic, clinical, pathologic, treatment, CTC this twenty year period were reported as MPM. Of the 6198 individuals grades of anemia and survival data were collected. A dedicated thoracic with a pleural sample during the twenty year period, 685 were diagnosed radiologist reviewed CT and PET-CT scans to assess pre-operative disease with MPM based upon the results of either electron microscopy performed distribution and post-operative recurrence. TTR and OS were calculated on the cytology specimen, or examination of a tissue biopsy specimen, from date of resection and estimated by the Kaplan-Meier analysis. Cox or necropsy. Of these 685 cases with pathologically confirmed MPM, 395 regression with dummy variables was used to assess the hazard ratios (HR) cases had a pleural cytology sample reviewed. From the table below, 283 associated with increasing CTC anemia grade relative to Grade 0 (HR = 1). of the 395 pathologically defined MPM cases (i.e. 72%) were diagnosed as MPM by cytopathologists in effusion specimens. Of the 112 patients with a Results: Median age was 63 years (range 27-86), and 182 (79%) patients biopsy diagnosis of MM with a cytology specimen which was not diagnostic were men. No patients received pre-operative chemotherapy (CT); 175 of MM; 49 were diagnosed as atypical or suspicious and 70 as negative (76%) received heated intra-operative chemotherapy (HIOC); 113 (49%) for malignancy. Of the cases not established by cytology, 40 (34%) were received adjuvant CT, and 75 (33%) received adjuvant radiation therapy sarcomatoid on biopsy. There were two incorrect diagnoses of MPM in (RT). For the 124 patients (54%) with Grade 0 anemia, median TTR and patients with metastatic malignancy to the pleura; both cases occurred OS were statistically significantly longer than for the 22 (10%) patients prior to availability of specific mesothelial markers. The absolute sensitivity with Grade 2 anemia. The TTR and OS rates were 18 and 21.8 months, of MPM effusion diagnosis was 78%; complete sensitivity was 87%, and respectively for patients with Grade 0 anemia compared with 3.2 and 6.4 the specificity was 99.9%. months, respectively for those with Grade 2 anemia. Similar statistically significant stratifications of TTR and OS by anemia grade were seen for each surgical procedure-- EPP and PD. Anemia was more prevalent in patients with non-epithelial histology, and occurred in 61% with non-

iMig2012.org • Abstract Book 50 epithelial histology compared with 38% of patients with epithelial histology. Anemia was associated with statistically significant decreased TTR and diminished OS for both histological subtypes. Details are shown in the table below.

CTC Anemia grade Median HR 95% CI Median OS months HR 95% CI TTR months All Patients 0 1 2 1.8 8.8 3.2 1.0 (1.5-3) 21.8 11.5 6.4 1.0 2.2 (1.6-3) 2.1 (2. 4-7) 3.5 (2.2-5.6) 4.1 EPP 0 1 2 18.8 9.6 3.3 1.0 (1.5-3.3) 20.2 11.8 7.3 1.0 1.7 (1.2-2.4) 2.2 (1.9-6.5) 2.5 (1.5-4.2) 3.6 september 2012 13, PD 0 1 2 14.7 7.7 3.8 1.0 (1.0-3.5) 42.8 11.4 6.0 1.0 5.0 (2.4-10.3) 1.9 - 16.9 (5.1-55.6) Epithelial 0 1 2 19.5 11.1 2.5 1.0 (1.3-3.0) 33.2 13.9 7.8 1.0 2.4 (1.6-3.6) Histology 1.9 (1.3-6.5) 3.3 (1.7-6.3) 2.9 Non-epithelial 0 1 2 15.4 7.6 5.4 1.0 (1.2-3.9) 15.2 9.1 6.0 1.0 1.5 (0.9-2.5) Histology 2.2 (1.9-10.0) 2.8 (1.3-5.9) 4.3

HR: Hazard Ratio; CI: Confidence Interval

Conclusion: Anemia is a clinical predictor for early recurrence and short overall survival independent of type of surgery or histology. Anemia can serve as a quantitative biomarker and should be included in the preoperative evaluation of MPM.

Disclosure: No significant relationships.

SESSION IVC pATHOLOGY September 13, 2012 14:20-16:00

IVC.6: WHOLE-SLIDE IMAGING DIGITAL PATHOLOGY AS A PLATFORM FOR ROUTINE MESOTHELIOMA SECOND LECTURE: A PILOT STUDY. EXPERIENCE OF THE MESOPATH CENTER.

Francoise Galateau Sallé1, Thomas Rousvoal2, Nolwenn Le Stang3, international mesothelioma interest group Issam Abdalsamad2, Hugues Begueret1, Elisabeth Brambilla1, Frederique Capron1, Marie Christine Copin1, Claire Danel1, Anne Yvonne De Lajartre1, Armelle Foulet Rogé1, Loulette Garbe1, Odile Groussard1, Christine Sagan1, Jean Michel Picquenot1, Francoise Thivolet Bejui1, Jean Michel Vignaud1, Jean Calude Pairon4, Patrick Brochard3, Anabelle Gilg Soit Ilg5 1Mesopath Centre, Chu Caen, Caen Cedex/FRANCE, 2Mesopath, Chu Caen, Caen/FRANCE, 3Pathology Department - University Hospital, Mesonat Registry (U 1086 Inserm), Caen Cedex 9/FRANCE, 4Inserm U955 Paris/ FRANCE, 5Invs/FRANCE

Background: Mesothelioma is a public health issue which exceeds the occupational framework and which presents major problems of pathological diagnosis, early detection towards effective therapy. In France, the collaborative effort of the domestic network MESOPATH and of the Center of Excellence IMEC made it possible to constitute a pole of research with international impact, in particular through a strong contribution to international programs for detection and classification of mesotheliomas. The clinico-biological database of MESOBANK a virtual and exhaustive repository of national data is interconnected with that of the Centre National Referent on pleural malignant mesothelioma and rare peritoneal tumors, supported by the INCA. Five thousands three hundred and ninety five cases have been validated from 1998 to 2011 according to a stringent procedure of certification previously described ( Goldberg et al, PNSM OEM,2006) with the disadventage of a long procedure. Whole-slide imaging technology (WSIT) offers promise for rapid, Internet- based telepathology consultations between institutions. To optimize the procedure of certification we evaluate the use of WSIT in mesothelioma pathological certification in respect to diagnostic accuracy, efficiency,

iMig2012.org • Abstract Book 51 clinical and research implications.

Methods: Slides (H&E and immunohistochemical slides including 2 positive and 2 negative markers for mesothelial cells) registered at the MESOPATH domestic referal center were scanned into whole digital format on a high quality LEICA virtual slide scanning system. Digital slides were independently diagnosed by 3 french panelists. 699 cases were scanned from June 1, to the 31 of May 2011. 5595 slides were examined by the pathologists through the CCITI application and retrieved on a local servor ( 2,69To). The results were compared to 844 cases certified according to the glass slides conventional certification. The diagnosis were performed through the CCITI software application. Reference, digital and glass-slide interpretations were compared. Operator comments on technical issues were gathered. september 2012 13, Results: Among the 699 cases, there was a 63% ( n=341) agreement on the diagnosis of mesothelioma among digital slides compared to a 79% ( n=670) results with conventional glass diagnoses. Minor disagreement were observed respectively in 30% ( n=162) with digital slides compared to 18% (n=153) and major disagreement ( benign versus malignant or sarcoma versus mesothelioma versus metastasis) in 7% ( n=38) compared to 3% ( n=27) with light microscope diagnoses. The minimal time for a complete expertise by three panelists with the digital slides system was 1h09 mn while the maximal time was 40 days compared to 30 to 100 days with the previous system using microscope analysis. The resolution was defined as fine by all panelists and the navigating ability at various magnification with the scanner and the CCITI application was well appreciated.

Conclusion: Our pilot study shows that dynamic whole-slides imaging is an excellent tool for second lecture and difficult cases such as those encountered with mesothelioma diagnosis. With further experience the accuracy of telepathology diagnosis will improve. It shows a termendous potential for rapid teleconsultation, research and education and as pathologist we may prefer in the future digital diagnostic practice.

Disclosure: No significant relationships.

SESSION IVC pATHOLOGY September 13, 2012 14:20-16:00

IVC.7: ROLE OF CYTOLOGY IN MESOTHELIOMA DIAGNOSIS international mesothelioma interest group Anders Hjerpe Laboratory Medicine, Karolinska Instiute, Stockholm/SWEDEN

Previous recommendations state that the diagnosis of malignant mesothelioma should be based on biopsy, while cytological diagnosis of exfoliated cells from an effusion is considered to be insufficient. However, the refinement of adjuvant techniques during the last decades has changed this completely. While the routinely stained cytological specimen may be inconclusive, numerous publications demonstrate the utility of effusion cytology in combination with ancillary techniques such as immunocytochemistry, biomarker analyses, FISH and/or electron microscopy. One such technique is often sufficient for diagnostic purposes. In a laboratory with experience of these techniques, mesothelioma diagnosis – when made - is accurate and definitive, and will in those cases provide all of the information necessary for choice of therapy. Diagnosis is usually obtained from the first effusion withdrawn, which in our material is 3-6 months before a biopsy is taken. This may save these patients from a series of cumbersome examinations. When the diagnosis is definite, additional biopsy sampling is unnecessary and may be considered unethical. I advocate that IMIG initiate a task force to define the conditions for accepting a mesothelioma diagnosis based on an effusion.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 52 Workshop VII Heated Chemotherapy and Other Approaches to Targeting Mesothelioma Surfaces September 13, 2012 17:00-18:30

WORKSHOP VII HEATED CHEMOTHERAPY AND OTHER APPROACHES TO tARGETING MESOTHELIOMA SURFACES September 13, 2012 17:00-18:30 september 2012 13,

WSVII.1: CHEMOTHERAPEUTIC OPTIONS FOR PERITONEAL AND PLEURAL MESOTHELIOMA, HIPEC, EPIC, HITAC AND HITOC – PHARMACOLOGIC STUDIES

Paul H. Sugarbaker Program In Peritoneal Surface Malignancy, Washington Cancer Institute, Washington, DC/UNITED STATES OF AMERICA

Background and Rationale: In the past peritoneal mesothelioma was treated by debulking surgery and systemic chemotherapy without demonstrable benefit. Over the past decade the median survival has been improved to over seven years.

Methods: Pharmacologic studies which establish the rationale for intraperitoneal and intravenous chemotherapy for this disease have been performed. These chemotherapy agents are now combined and appropriately administered either intravenously or intraperitoneally in an attempt to maximize responses. The chemotherapy agents are used perioperatively.

Results: Currently, the chemotherapy is given into the peritoneal cavity with heat (HIPEC) into both peritoneal and pleural cavities simultaneously (HITAC) or used within the pleural space only (HITOC). Chemotherapy clearance from the abdomen is more rapid than from the pleural space. Pharmacologic studies of intraperitoneal doxorubicin, intraperitoneal cisplatin, systemic ifosfamide, intraperitoneal paclitaxel, intraperitoneal pemetrexed, intraperitoneal gemcitabine, and intraperitoneal mitomycin

C are to be illustrated. Multi-agent chemotherapy by intraperitoneal and international mesothelioma interest group intravenous routes of administration with acceptable toxicity are the goals of this effort.

Conclusions: Bidirectional chemotherapy can be used to improve the results of treatment with peritoneal mesothelioma at three different time periods. Currently, we utilize a five-drug protocol for comprehensive management. The chemotherapy treatments are used as a planned part of the combined surgical and chemotherapeutic treatment plan for these patients. Comprehensive morbidity/mortality assessments accompany all management plans.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 53 General Session V Asbestos and Mesothelioma September 14, 2012 08:00-09:30

GENERAL SESSION V ASBESTOS AND MESOTHELIOMA 1.64 and 1.78 for 2006-9. This is well under half of most recently reported September 14, 2012 08:00-09:30 rates in France, Italy, and especially the UK and Australia. (It should be

noted of course that while this is good news, the raw number of cases september 14, 2012 GSV.1: UPDATE ON EPIDEMIOLOGY IN NORTH AMERICA annually in the US remains higher than in any other country, and this is likely to remain true for many years even as rates continue to decline). Bruce W. Case An important and controversial issue relates to the health effects of Pathology Epidemiology And Occupational Health, McGill University, nonasbestiform amphibole varieties (in several mineralogical categories) . Montreal/CANADA Although a great deal has been written (particularly about talc, tremolite, and taconite) except for ecological observations (Case BW et al. J Toxicol This brief presentation is an “update” of some findings and trends in Environ Health B Crit Rev. 2011;14(1-4):3-39) this is a difficult area to the epidemiology of mesothelioma in the United States and Canada. A study epidemiologically. An extensive Taconite Workers Health Study PubMed search for English papers published 2009-2012 containing the being performed at the University of Minnesota is one such case (http:// words mesothelioma and epidemiology produced 302 papers; about taconiteworkers.umn.edu/index.html) ; amphiboles are present but there half were from the USA or Canada. One Mexican case-control study is debate about their nature; mesothelioma SMR increase and a high rate of 472 workers (117 mesothelioma cases and 353 controls) insured by of radiological abnormalities is reported as present by the investigators to the Mexican Institute of Social Security (Aguilar-Madrid et al. 2010, date but incidence, analysis and other studies are not yet complete, and a Am J Indust Med) had qualitative exposure assessment which indicated previous study suggested that cases among workers had sources of (prior) “overall proportion of certain, likely, and possible occupational asbestos exposure other than the mining dust itself. A registry of the former workers exposure in some workplaces with exposure of asbestos” as 80.6% in of the Baie Verte asbestos mine in Newfoundland has been developed cases and 31.5% in controls). Historical occupational cohort studies by SafetyNet at Memorial University in St.-John’s with the cooperation of (e.g. chrysotile miners and millers of Quebec; textile workers in the former workers. Information on 1003 voluntary registrants (denominator Carolinas; the W.R. Grace vermiculite mining and processing cohorts) uncertain; some 2400 to 3000 ever worked) has been obtained for health saw one new group (North Carolina textile) and some follow-ups. Most status, disease outcomes, and cumulative exposure using a JEM procedure recent work is methodological rather than new case-finding work. New with access to over 8000 historical air-samples from company, union and case-control studies are rare and add little to what was already known, government sources. Results for this chrysotile mine are expected in 2012. although one (Pintos et al., J Occ Env Med 2009) suggested a possible Finally, after three decades of study by many groups, EPA is in the final interaction between asbestos and “man-made” mineral fibers. Arguably stages of carrying their assessment of exposure and disease in the W.R. more important new epidemiological work was done during this period Grace vermiculite mining workforce at Libby, Montana and related sites in Italy, France, and the United Kingdom. Nevertheless some new work (Toxicological Review of Libby Amphibole Asbestos in Support of Summary has been accomplished and some important work will be reported within Information on the Integrated Risk Information System (IRIS)) forward to the next six months. The methodological papers include a debate about use in the IRIS database. This is being done through the efforts of a Review

the type of data suitable for risk analysis, with two roads to the same Panel set up under the auspices of EPA’s Science Advisory Board. Drafts international mesothelioma interest group goal - hierarchical ranking of studies with sequential exclusions versus of the Panel Report are available on the EPA website; the most recent use of all available data with adjustments for various quality elements and Deliberative Draft at this writing is at http://yosemite.epa.gov/sab/ uncertainty (e.g. Berman DW and Case BW; Ann Occup Hyg (2012) doi: sabproduct.nsf/ea5d9a9b55cc319285256cbd005a472e/e9214b4949ef0f 10.1093/annhyg/mes027 First published online: July 23, 2012). In Canada, e985257a380067c4e7/$FILE/Libby%20Asbestos%20Report%207-11-12. studies using administrative data from four provinces have explored the pdf. Considerable public comment and comments from EPA and from the extent and nature of mismatches between registered mesothelioma cases panelists themselves is available via the EPA SAB websites. and compensated mesothelioma cases. Studies from Quebec, Alberta, and most recently British Columbia and Ontario have all shown far lower Disclosure: No significant relationships. percentages of registered cases receiving compensation than would be expected, even though in these jurisdictions almost all claims with disease are accepted. Under-reporting to the compensation boards or failure to make claims is clearly responsible but the reasons are unclear. Making GENERAL SESSION V ASBESTOS AND MESOTHELIOMA reporting legally mandatory within 14 days of diagnosis (in Quebec) and September 14, 2012 08:00-09:30 increased communication between registries and compensation boards have produced some improvement. A recent study (Labrèche F et al. Can GSV.2: MESOTHELIOMA AND THE LAW Respir J. 2012 19:103-7) demonstrates that the low compensation rate in Quebec is not due to over-registration since 62 to 77% of registered David Rosenberg cases were confirmed probable or definite and an additional ten to 19% Harvard Law School, Boston/UNITED STATES OF AMERICA possible. In fact, separate study of compensated cases showed that 13 per cent had been misclassified in the tumour registry as other cancer Exposure of millions of workers to asbestos has resulted in the largest, types, although true total underascertainment could not be evaluated. longest running, and most expensive mass tort litigation in U.S. history. Incidence trends will be discussed: an important observation is the now Judge by its main justification – compensation – asbestos litigation well established year-over-year decrease in male cases in the United could properly be labeled a fiasco were it not for its gigantic waste of States which has seen a significant Average Annual Percent Change of -1.4 social resources. Well over $70 billion has been spent since the 1970s on to -1.6 per cent per year over the last ten years (2000-2009) (National overhead – lawyers, experts, insurance and court administration, and Cancer Institute; SEER Cancer Statistics Review 1975-2009; last updated other litigation related matters – while $30 billion in net compensation August 20 2012; http://seer.cancer.gov/csr/1975 _ 2009 _ pops09/ has thus far been received by plaintiffs. This cost-benefit ratio translates results _ merged/sect _ 17 _ mesothelioma.pdf). Overall, age- into more than $1.40 being spent to deliver $1.00 in compensation to adjusted incidence from the SEER 9 data set for males peaked at 2.17 per each plaintiff (who probably suffered little or no disabling injury and likely 100,000 population in 1994 and 1995 and most recently was between was never exposed to asbestos). As such, asbestos litigation is more

iMig2012.org • Abstract Book 54 accurately characterized as antisocial. Nearly all of this colossal profligacy could have been avoided. Contrary to the prevailing view, the solution is not creation of a 9-11 or BP Oil Spill type fund. All the courts should have done (and need to do in the future for asbestos and other mass tort cases) is to pay strict attention to insurance theory and to forgo pursuit of the compensation goal entirely.

Disclosure: No significant relationships. september 14, 2012 international mesothelioma interest group

iMig2012.org • Abstract Book 55 Session VA1 Peritoneal Mesothelioma September 14, 2012 10:00-11:00

SESSION VA1 pERITONEAL MESOTHELIOMA In an analysis of these experiences, median survival increased from 12 September 14, 2012 10:00-11:00 months with systemic chemotherapy treatment to 53 months with CRS and

HIPEC, with a 50% 5 year overall survival. september 14, 2012 VA1.1: PERITONEAL MESOTHELIOMA: INTEGRATION OF MULTIMODAL TREATMENT AND TRANSLATIONAL RESEARCH Molecular biology During the last decade, DMPM biology has been deeply explored at the Milan NCI through various clinical biological studies. Marcello Deraco1, Nadia Zaffaroni2, Federica Perrone3 Telomerase activity (TA) was recently reported to be expressed in the 1Surgery, Fondazione Irccs Istituto Nazionale Tumori, Milano/ majority of DMPMs and to negatively affect the clinical outcome of DMPM ITALY, 2Department Of Experimental Oncology, Irccs Istituto Nazionale patients (Villa et al). In a study, 44 DMPM specimens were analyzed; TA Tumori, /ITALY, 3Pathology, Irccs Isituto Nazionale Tumori Milano/ITALY was determined using the telomeric repeat amplification protocol, and alternative-lengthening of telomeres (ALT) was detected by assaying ALT Recent clinical and basic science research efforts have changed the associated promyelocytic leukemia nuclear bodies. In the overall series, perception of diffuse malignant peritoneal mesothelioma (DMPM). This TA was prognostic for 4-year relapse and cancer related death, whereas clinical entity presents epidemiologic, biological, and clinical behaviors that ALT failed to significantly affect clinical outcome. These results held true are different from its more frequent pleural counterpart. A multimodality in the subset of patients who underwent uniform treatment with CRS treatment consisting of cytoreductive surgery (CRS) and hyperthermic and HIPEC. Zaffaroni et al investigated the proliferation and apoptotic intraperitoneal chemotherapy (HIPEC) has emerged as the most effective features of DMPM by assessing the immunohistochemical expression of approach for the treatment of DMPM. In addition, new therapeutic targets survivin and members of the IAP family, including IAP 1, IAP 2, and X-IAP, and prognostic biological factors have been identified. The incidence of in a series of 32 DMPM specimens. The effects of survivin knockdown DMPM corresponds to approximately one fourth of pleural mesothelioma in an established DMPM cell-line were also appraised. DMPM cells were and has been rising worldwide since 1970. The annual mortality rate transfected with small interfering RNA (siRNA) targeting survivin mRNA and is expected to increase 10% worldwide until 2020. Age-standardized analyzed for survivin expression, growth rate, and the ability to undergo incidence rates among men range from 0.5 to 3 cases/1,000,000. spontaneous and chemotherapy induced apoptosis. DMPM cells were proven to be characterized by a relatively low proliferative activity and Systemic Therapies DMPM is known to be chemoresistant and prolonged the low apoptotic behavior. Moreover, survivin and other IAPs are found survival resulting from systemic chemotherapy has not been reported yet. to be largely expressed in DMPMs and suggest that strategies aimed at Numerous single drug and combination regimens have been tested over down regulating survivin (transfection of DMPM cells with survivin siRNA) the past decades with modest results. A systematic review (2002) including may provide a novel approach for the treatment of this malignancy. The more than 2,300 patients (pleural and peritoneal combined) in 83 trials. authors have also assessed the cellular effects of new anticancer agents, Regarding antitumoral response rate, cisplatin was suggested to be the including a new series of nortopsentin hetero analogues in a DMPM cell- most active single agent, and the combination of cisplatin+doxorubicin, the line. Selected compounds that were able to inhibit the activity of CDK1 most active regimen. Data from a German study and from the pemetrexed consistently reduced cell growth and induced a concentration dependent international mesothelioma interest group expanded access program on the antitumor activity of pemetrexed cell cycle arrest at the G2/M phase and an increase in the apoptotic rate +/-platinum in DMPM suggested response rates in the range of those with a concomitant down regulation of the anti apoptotic protein survivin. observed for pleural disease. Preliminary data suggested a possible Notably, the addition of these compounds to paclitaxel treated cells survival advantage for the combination of cisplatin+raltitrexed compared resulted in a marked increase of the cytotoxic effect as a consequence with cisplatin alone. Other chemotherapies that have been shown to be of increased caspase activation. Recently, Pilotti and Perrone aimed active in this setting include vinorelbine and gemcitabine, either alone to assess the activation profile of EGFR, PDGFRB and PDGFRA receptor or in combination with platinum compounds. In historical case series, tyrosine kinases (RTK) and their downstream effectors in cryopreserved standard therapy with palliative surgery and systemic or intraperitoneal DMPM specimens. They also made a complementary analysis of the chemotherapy has been associated with a median survival of about one cytotoxic effects of some kinase inhibitors on the proliferation of the year (9 to 15 months). human peritoneal mesothelioma STO cell-line. They found the expression/ phosphorylation of EGFR and PDGFRB in most of the tumours, and PDGFRA CRS and HIPEC DMPM is generally confined to the peritoneal cavity activation in half. The expression of the cognate ligands TGF α, PDGFB and rarely metastasizes systemically. Most patients die because of and PDGFA in the absence of RTK mutation and amplification suggested complications directly related to intra abdominal disease progression, the presence of an autocrine/paracrine loop. There was also evidence such as bowel obstruction and starvation. CRS, through peritonectomy of EGFR and PDGFRB co-activation. RTK downstream signaling analysis procedures and multivisceral resections, allows the removal of all visible demonstrated the activation/expression of ERK1/2, AKT and mTOR, S6, tumor implants, and HIPEC, through intra-abdominal drug administration and 4EBP1, in almost all the DMPMs. No KRAS/BRAF mutations, PI3KCA with hyperthermia, is used to treat the free tumor cells and very small mutations/amplifications or PTEN inactivation were observed. Real- residual disease. At the National Cancer Institute (NCI) of Milan, CRS is time PCR revealed the decreased expression of TSC1 c DNA in half of the performed according to the technique originally described by Sugarbaker tumours. In vitro cytotoxicity studies showed the STO cell-line to be with some minor technical variants. HIPEC is performed with the closed resistant to gefitinib and sensitive to sequential treatment with RAD001 abdomen technique with cisplatin (45 mg/L of perfusate) and doxorubicin and sorafenib; these findings were consistent with the presence of the (15 mg/L of perfusate) for 90 minutes at a temperature of 42.5oC. KRAS mutation G12D in these cells although it was not detectable in the Perfusate volume was 4 to 6 L, and mean flow was 700 mL/min and the original tumour. extracorporeal circulation device Performer LRT® (RAND, Medolla, Italy) is used. Our center has gathered more than 15 year experience with an Future perspectives Taking advantage of available array based Institutional casuistic of more than 150 cases operated up to now. The approaches, studies aimed at evaluating gene, microRNA and protein combined approach of CRS with HIPEC modified the natural history of expression in a high throughput fashion are currently ongoing to elucidate DMPM, introducing a dramatic improvement in outcomes in principal the biological mechanisms and the regulatory events responsible for DMPM dedicated international centers and in a multi-institutional registry series. incidence and progression. Results from these studies are expected to pave

iMig2012.org • Abstract Book 56 the way for the identification of novel therapeutic targets and strategies as well as new biomarkers for the selection of patients who could benefit from specific treatments.

Disclosure: No significant relationships.

SESSION VA1 pERITONEAL MESOTHELIOMA September 14, 2012 10:00-11:00

VA1.2: DIFFUSE MALIGNANT PERITONEAL MESOTHELIOMA: SEVEN-YEAR ACTUAL SURVIVAL AFTER SURGICAL CYTOREDUCTION AND HYPERTHERMIC INTRAPERITONEAL september 14, 2012 CHEMOTHERAPY (HIPEC) DEFINES CURE.

Dario Baratti1, Shigeki Kusamura1, Antonello D. Cabras2, Rossella Bertulli3, Marcello Deraco1 1Surgery, Fondazione Irccs Istituto Nazionale Tumori, Milano/ ITALY, 2Pathology, Fondazione Irccs Istituto Nazionale Tumori, Milano/ ITALY, 3Adult Mesenchymal Tumor Medical Oncology Unit, Fondazione Irccs Istituto Nazionale Tumori, Milano/ITALY Conclusion: Patients with DMPM who survived ≥7 years appeared to be cured by CRS and HIPEC. Cure rate was 45.8%. Proliferative index may be Background: Diffuse malignant peritoneal mesothelioma (DMPM) is a rare, used for prognostic stratification of DMPM. Biological features, such as locally aggressive, and rapidly lethal neoplasm. Cytoreductive surgery CK5/6, EGFR, and podoplanin, warrant further investigations. (CRS) combined with hyperthermic intraperitoneal chemotherapy (HIPEC) is an innovative treatment option for peritoneal surface malignancies. Disclosure: No significant relationships. Because of the current lack of prospective randomized data, a survival benefit for DMPM patients managed by the local-regional approach has been suggested by comparison with historical controls, based on actuarial data. The aims of this study were to define cure after combined treatment SESSION VA1 pERITONEAL MESOTHELIOMA for DMPM, determine the cure rate based on actual survivors, and identify September 14, 2012 10:00-11:00 clinical-biological features associated with cure. VA1.3: ADJUVANT BIDIRECTIONAL CHEMOTHERAPY Methods: A prospective database of 132 patients with DMPM undergoing WITH INTRAPERITONEAL PEMETREXED COMBINED WITH CRS and HIPEC was reviewed. CRS comprised peritonectomy procedures INTRAVENOUS CISPLATIN FOR DIFFUSE MALIGNANT PERITONEAL and visceral resections, as needed. Close-abdomen HIPEC was MESOTHELIOMA: PHARMACOLOGIC AND PHASE II STUDY performed with cisplatin and doxorubicin or mitomycin-C. A panel of immunohistochemical markers related to the origin and peculiar clinical Paul H. Sugarbaker1, O Anthony Stuart2, Lana Bijelic3 features of DMPM was tested. Cell proliferation was scored by the 1Program In Peritoneal Surface Malignancy, Washington Cancer Institute, percentage of tumor cells expressing the Ki-67 nuclear antigen. Washington, DC/UNITED STATES OF AMERICA, 2Medstar Research Institute, Washington, DC/UNITED STATES OF AMERICA, 3Department Of Surgery, Results: Operative mortality occurred in 3 patients (2.2%) and major Washington Hospital Center, Washington, DC/UNITED STATES OF AMERICA international mesothelioma interest group morbidity in 46 (34.3%). In the overall series, median follow-up was 54.8 months (95% confidence interval (CI) 43.2-66.5). Median actuarial Background: Cytoreductive surgery (CRS) and heated perioperative survival was 71.9 months (95%CI 32.3-111.5), and actuarial 5-year chemotherapy (HIPEC) have successfully moved the median survival of survival was 55.6%. The survival curve reached a plateau at 7 years, diffuse malignant peritoneal mesothelioma (DMPM) from 12 to 60 months representing 22 actual survivors of 48 patients (45.8%) who had the and is now considered a standard of care. Nevertheless despite careful potential for at least 7 years of follow-up (see figure). Among these patient selection for CRS and HIPEC progressive disease does occur. 7-year survivors, 19 patients were free of disease and only one patient Adjuvant treatments for DMPM have not been developed and need to be experienced disease-specific death after 7 years. Cytokeratin 5/6, investigated. We prospectively studied intraperitoneal pemetrexed and calretinin, Wilms Tumor-1 (WT-1), podoplanin, epithelial growth factor intravenous cisplatin as an adjuvant treatment. receptor (EGFR), matrix metalloproteinase-2 were mostly positive; p16, and matrix metalloproteinase-9 were poorly expressed. At multivariate Methods: From September 2007 until September 2009, 10 patients with analysis, completeness of cytoreduction (P=0.002), histological subtype DMPM were enrolled in a pharmacologic and phase II, single-institution (P=0.025), no visceral resections (P=0.012), and ≤10% Ki67-positive cells protocol of adjuvant intraperitoneal (IP) pemetrexed combined with (P=0.004) independently correlated with long-term survival, although no intravenous (IV) cisplatin following best surgical cytoreduction. The preoperative clinical-biological factor was sufficiently discriminating to treatments were given every 3 weeks for 6 cycles and consisted of 500 preclude cure. At univariate analysis, increased progression-free survival mg/m2 of IP pemetrexed and 75 mg/m2 of IV cisplatin. Pemetrexed was correlated with negative/weak EGFR (P=0.049), positive CK5/6 (P=0.025), delivered through a peritoneal port placed at the time of cytoreduction. and positive podoplanin (P=0.001). After multivariate analysis, only positive Toxicities were prospectively evaluated using the Common Toxicity Criteria, podoplanin retained its prognostic significance for better progression-free Version 2.0. survival (P=0.001). Results: The mean age of the patients was 51 years and 7 of 10 were male. Eight patients had epithelioid mesothelioma and 2 had biphasic. The mean peritoneal cancer index (PCI) was 24. Four patients had a CC-1 cytoreduction, 4 had CC-2 and 2 had CC-3. Two patients had a colon resection as part of their surgery. Nine of the 10 patients were able to complete all 6 cycles of therapy. One patient had a catheter infection requiring catheter removal and completed two cycles of intravenous therapy. No patients required dosing modifications. Five of the 10 patients (50%) experienced grade I nausea. Four patients (40%) experienced grade I and one patient (10%) grade II abdominal pain. There were no hematologic

iMig2012.org • Abstract Book 57 toxicities and no grade III or IV toxicities except the catheter infection. Chromosomal aberrations were compared statistically between the Pharmacologic analysis of pemetrexed peritoneal and plasma levels was asbestos and radiation groups and one prominent region was evidenced in performed showing a peritoneal fluid to plasma AUC ratio of 70. After a chromosome 14q where significant loss was shown in asbestos cases, but median follow-up of 60 months, the median survival of these 10 patients not radiation cases as highlighted in Figure 1. has not been reached; 3 patients have no evidence of disease, 3 are alive with disease and 4 have died of disease. Conclusion: Copy number analysis using SNP mapping arrays revealed losses at 14q11.2-13.3 and 14q21.1-23.2 in up to 70% of PMM patients after Conclusion: Adjuvant intraperitoneal pemetrexed combined with asbestos exposure compared to no deletion in PMM patients after radiation intravenous cisplatin following cytoreduction for DMPM can be exposure. While the significance of this loss is unknown, identification in administered with very morbidity and has favorable short-term outcomes. asbestos induced PMM in contrast to radiation attributed PMM suggests a The intraperitoneal administration of pemetrexed has important distinct molecular pathway for asbestos induced versus radiation induced pharmacologic advantages and is associated with few adverse events. epithelioid PMM.

Disclosure: No significant relationships. Disclosure: No significant relationships. september 14, 2012

SESSION VA1 pERITONEAL MESOTHELIOMA September 14, 2012 10:00-11:00

VA1.4: IDENTIFICATION OF CHROMOSOME 14Q DELETIONS IN ASBESTOS-INDUCED AND NOT RADIATION-INDUCED PERITONEAL MALIGNANT MESOTHELIOMA

Katherine Chen1, Robert N. Taub2, Joseph Testa3, Jianming Pei4, Brynn Levy1, Odelia Nahum1, Jin Li Chen1, Adam Zimilover1, Alain Borczuk1 1Herbert Irving Comprehensive Center, Columbia University, Washington/ UNITED STATES OF AMERICA, 2New York Presbyterian Hospital/UNITED STATES OF AMERICA, 3Fox Chase Cancer Center Of Temple University School Of Medicine, Philadelphia/UNITED STATES OF AMERICA, 4Fox Chase Cancer Center Of Temple University School Of Medicine/UNITED STATES OF AMERICA

Background: Malignant mesothelioma is an aggressive tumor arising from mesothelial lining of serosal cavities. Pleural is the most common site, accounting for 60-70% of mesothelioma cases, and peritoneum accounts for the majority of the remaining 30%. Asbestos exposure is associated with pleural mesothelioma, with an attribution of 70%. Peritoneal malignant mesothelioma (PMM), in contrast, has a weaker attribution to asbestos of 30%. Other recognized causal factors for peritoneal mesothelioma include radiation therapy following testicular cancer, breast cancer, and lymphoma. The disease appears to be similar regardless of causation, but the molecular pathogenesis of asbestos versus radiation attributed peritoneal mesothelioma is unknown. international mesothelioma interest group

Methods: We identified 37 cases of snap frozen epithelioid PMM tissue from the Cancer Center Tissue Bank. Manual microscope guided needle dissection of cryostat frozen sections was performed obtaining a minimum of 500ng of tumoral DNA. DNA was prepared as per Affymetrix protocol for digestion, ligation, amplification, purification and labeling with standard quality metrics for fragment size prior to hybridization on Affymetrix SNP 6.0 array. Array results were analyzed using Nexus Copy Number 6.0 software (Biodiscovery, Hawthorne CA).

Results: Ten of the 37 PMM cases were attributed to asbestos exposure and seven to radiation exposure. In asbestos attributed cases, patient age ranged from 45-78 (8 men and 2 women) and in the radiation attributed cases, patient age ranged from 49-88 (6 men and 1 woman).

Figure 1. Frequency of chromosome loss and gain in asbestos (red) versus radiation (blue) induced PMM When chromosome 14q was further analyzed, loss of 14q11.2-14q13.3 and 14q21.1-23.2 was seen in up to 7 of 10 cases (70%) of asbestos attributed mesothelioma and in 0 of 7 (0%) of the radiation attributed cases.

iMig2012.org • Abstract Book 58 Session VA2 Asbestos/Nanoparticles September 14, 2012 11:00-11:30

SESSION VA2 ASBESTOS/NANOPARTICLES September 14, 2012 11:00-11:30 september 14, 2012 VA2.2: LENGTH- AND TIME-DEPENDENT CELLULAR SIGNALLING RESPONSE TO ASBESTOS AND CARBON NANOTUBES EXPOSURE.

Tatyana Chernova1, Anja Schinwald2, Fiona Murphy1, Xiao Ming Sun1, Martin Bushell1, Ken Donaldson2, Anne E. Willis1, Marion Macfarlane1 1Toxicology Unit, Mrc, Leicester/UNITED KINGDOM, 2Mrc/University Of Edinburgh, Centre For Inflammation Research, Qmri, Edinburgh/UNITED KINGDOM

Background: Exposure to asbestos fibres causes profound pathological changes and can result in the development of a fatal tumour, malignant mesothelioma (MM). There is rising concern that carbon nanotubes (CNTs) may present an inhalation hazard similar to asbestos and cause MM.

Methods: To investigate the molecular changes that occur as a consequence of direct exposure to short and long asbestos fibres (SFA and LFA) and short and long CNTs (NTS and NTL) these types of fibres were instilled directly into the pleural cavity in mice, the site of mesothelioma development, and kinome profiling was performed after 1, 4 and 12 weeks of treatment.

Results: Exposure to fibres resulted in both acute and delayed responses that were dependent upon the length of the fibres and the time of exposure. In response to NTL and LFA there was acute inflammation and fibrosis on the parietal pleura; no inflammatory changes were detected histologically after exposure to SFA and NTS. In terms of changes to the kinome, exposure to SFA and NTS produced only an acute response after 1 week, which involved activation of MSK1/2, Akt, RSK1/2/3, p53 and

p27. The changes observed in LFA and NTL were similar, but in addition, international mesothelioma interest group there was sustained activation (during the entire length of the treatment) of mTOR, Src family kinases and STAT3, failure to activate p27 and p53, and reduced STAT-1 activation. Importantly, parallel profiling of malignant mesothelioma tumour samples identified similar changes in these signalling pathways.

Conclusion: The contribution of different cell types in time-dependent activation of these signalling pathways their role in disease development will be discussed.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 59 Session VB Molecular Therapy and Genomics September 14, 2012 10:00-11:30

SESSION VB mOLECULAR THERAPY AND GENOMICS exposure appears to be necessary in most cases for the development September 14, 2012 10:00-11:30 of mesothelioma, it is insufficient alone to cause mesothelioma. This

is inferred by the fact that nearly 27 million individuals in the US were september 14, 2012 VB.2: IDENTIFYING THERAPEUTIC TARGETS FOR MESOTHELIOMA exposed to asbestos in the work place between 1940 and 1979 but just USING SIRNA 3,000 new cases of mesothelioma are diagnosed annually. Common cancers are 3 times more likely to occur in mesothelioma patients and their Cleo Robinson1, Elliot Sollis2, Ian Dick2, Jenette Creaney1, Richard Lake2 1st degree relatives. Hence genetic susceptibility may affect mesothelioma 1School Of Medicine And Pharmacology, National Centre For Asbestos development. Furthermore, it seems likely that the genetic signature Related Diseases, University Of Western Australia, Perth/WA/ of asbestos induced mesothelioma may differ from that of peritoneal AUSTRALIA, 2School Of Medicine And Pharmacology, Ncard, University Of mesothelioma that affects women without known asbestos exposure. Western Australia, Perth/WA/AUSTRALIA Methods: A genome-wide association study (GWAS) was performed on Background: Mesothelioma is essentially incurable and new drugs to 203 individuals with mesothelioma genotyped with the CNV370 BeadChip effectively treat it are urgently needed. Our strategy to achieve this aim (Illumina, Inc.). Control subjects for this population were an independent was to identify candidate mouse and human genes that may have a role group of Caucasian individuals from the iControls dataset (https://www. in mesothelioma development and to inhibit their expression in fully illumina.com/science/icontrodb.ilmn). The 3172 iControl subjects were transformed mesothelioma cell lines using siRNA. genotyped using Illumina Hap300 and Hap550 Chips. For both case and control population, we imputed all the known SNPs using IMPUTE program. Methods: Selection of genes was on the basis of their differential Individuals with a call rate <0.95 were removed from GWAS analysis. The expression in transcriptome or CGH analyses when comparing malignant to following quality control criteria were used to filter SNPs: Minor Allele normal mesothelial cells, together with any known functional information Frequency < 0.02, Hardy-Weinberg Equilibrium < 0.001 and call rate which could be relevant to tumorigenesis. In a similar way we also selected <0.95. After imputation and applying quality control, there were 195 a small number of candidates from other published studies. A second cases and 2847 controls with 2,016,892 common SNPs. The association set of candidates was chosen from aberrantly expressed kinases with testing was first performed using all 195 mesothelioma cases. Twelve the idea that these genes are more likely to represent druggable targets female subjects with peritoneal mesothelioma were then removed from the given the broad range of kinase inhibitors that are widely available. dataset and the association tests were performed again; both using linear Where possible, we identified mouse and human homologues of the 40 regression implemented in PLINK. Population stratification was adjusted by candidates and then generated both mouse and human siRNA libraries. We the top 2 eigenvectors using EIGENSOFT software (Price et al.2006). tested the effect of gene knockdown on the growth of mouse and human mesothelioma cell lines in vitro. Results: Six SNPs (rs16872571, rs2059109, rs4505994, rs7765557, rs7817028 and rs1491485) were significant (p < 10-8)after Bonferroni

Results: We found knockdown was efficient and inhibition of a subset correction comparing cases to Illumina controls. Four of these 6 SNPs international mesothelioma interest group of the selected genes slowed cell growth significantly across a range of have been shown to be associated with common cancers. rs2059109 cell lines in both mouse and human systems. There was not complete (CCNG2) is a tumor suppressor gene that is involved in regulation of concordance between the mouse and human: Incenp, Plk1 and Tpx2 telomerase activity, rs7765557 (CD109) and rs7817028 (RUNX1T1) interact were found to be important for murine cellular proliferation; whereas, with and negatively modulate TGF-B signaling and rs1491485 (FAM84B) AURKA, TPX2 and BIRC5 were found to be important for human cellular through unknown mechanisms. rs16872571 (CLNK also known as MIST) proliferation. KIF11 was identified in both studies. regulates NK cell cytotoxicity. No known association has been reported with rs4505994. The same 6 SNPs were also significant after removing Conclusion: These genes all have a function in chromosome positioning, the 12 women with peritoneal mesothelioma but the p values became less centrosome separation and spindle assembly during cell mitosis. We significant. There were however, 3 regions where an improved signal was will present data to show whether they have an additional role in cell seen after removal of the women with peritoneal mesothelioma (chr9, migration, cell invasion and apoptosis. We think these studies could chr11, chr17). provide new leads for drug development. Conclusion: Six SNPs were identified that were associated with Disclosure: No significant relationships. susceptibility to mesothelioma. Removal of women with peritoneal mesothelioma had little impact on this association. Association of peritoneal mesothelioma in women with regions in chr9, chr11, chr17 require further investigation. SESSION VB mOLECULAR THERAPY AND GENOMICS September 14, 2012 10:00-11:30 Disclosure: No significant relationships. VB.3: GENOME WIDE ASSOCIATION STUDY IN MESOTHELIOMA COMPARED TO ILLUMINA CONTROLS SESSION VB mOLECULAR THERAPY AND GENOMICS Jill Ohar, Sha Tao, Tim Howard, Jianfeng Xu September 14, 2012 10:00-11:30 Genomics, Wake Forest University, Winston Salem/NC/UNITED STATES OF AMERICA VB.4: MOLECULAR CLASSIFICATION OF HUMAN MALIGNANT PLEURAL MESOTHELIOMA Background: A history of asbestos exposure can be identified in more than 80% of mesothelioma victims. However, women with peritoneal Didier Jean1, Aurélien De Reynies2, Annie Renier1, Ilir Hysi1, Françoise mesothelioma often lack an asbestos exposure history. Though asbestos Le Pimpec-Barthes3, Pascal Andujar4, Marie-Christine Copin5, Paul

iMig2012.org • Abstract Book 60 Hofman6, Jean-claude Pairon7, Françoise Galateau-Sallé8, Jessica Results: Gene expression profiling by microarray of MPM in culture Zucman-Rossi1, Marie-Claude Jaurand1 allowed to define two main clusters (C1 and C2), and a third cluster 1Umr-S 674, Inserm - Université Paris Descartes, Paris/FRANCE, 2Ligue of normal mesothelial cells. We demonstrated that mRNA expression Nationale Contre Le Cancer, Paris/FRANCE, 3Service De Chirurgie analysis by RT-qPCR using the most discriminating genes allowed the Thoracique, Hôpital Européen Georges Pompidou, Assistance Publique same clusterization of MPM in culture, and divided MPM tumor samples - Hôpitaux De Paris, Paris/FRANCE, 4Service De Pneumologie Et De into two clusters included into C1 and C2 respectively. In MPM in culture, Pathologie Professionnelle, Centre Hospitalier Intercommunal De mutations in CDKN2A, CDKN2B, NF2 and BAP1 are more frequent in cluster Créteil, Créteil/FRANCE, 5Pôle Biologie Pathologie, Chru De Lille, Lille/ C1 than in cluster C2; the difference is statistically significant forBAP1. FRANCE, 6Laboratoire De Pathologie Clinique Et Expérimentale, Chu De This classification also distinguished MPM according to their histological Nice, Nice/FRANCE, 7Service De Pneumologie Et Pathologie Professionnelle, subtypes for both MPM in culture and tumor samples. Sarcomatoid and Centre Hospitalier Intercommunal De Créteil, Créteil/FRANCE, 8Service desmoplastic MPM are only found in cluster C2. Interestingly, we observed D’Anatomie Pathologique, Chu De Caen, Caen/FRANCE a longer patients’ survival in clusters C1 compared to C2. An analysis of more tumor samples is underway to confirm this observation. Background: Malignant Pleural Mesothelioma (MPM) classification is based on histology, and there is no definitive classification based on molecular Conclusion: This classification allows to define a new subclass of september 14, 2012 profile, which take into account the biological diversity of this tumor. epithelioid MPM, which clusterized with sarcomatoid MPM. MPM of the Considering MPM molecular features should allow designing targeted two clusters have different prognosis and, surprisingly, C1 cluster with therapies, a strategy previously shown to be beneficial in other cancers the highest mutation frequency in tumor genes is characterized by the such as lung or breast carcinomas. The objective of this study was to highest median survival. Our data also showed several signaling pathways establish a MPM molecular classification of therapeutic, diagnostic and differently regulated between the two clusters. Molecular markers for prognostic interests. epithelial-mesenchymal transition have a higher expression in MPM in cluster C2. Methods: We performed a large-scale transcriptomic analysis on 38 MPM and 3 normal mesothelial cells in primary culture by microarray analysis Disclosure: No significant relationships. and defined MPM clusters by unsupervised hierarchical classification (CIT program/LNCC). Molecular clusterization was validated on a higher number of MPM in culture (67 cases) by studying mRNA expression of 87 genes by RT-qPCR using TLDA technology. In parallel, 57 MPM tumor samples were also analyzed to evaluate this classification in tumor samples. The phenotype of each cluster was characterized by taking into account clinical and epidemiological (asbestos exposure) data and the mutation status of several genes involved in mesothelial carcinogenesis (CDKN2A, CDKN2B, NF2, BAP1, TP53 and CTNNB1). international mesothelioma interest group

iMig2012.org • Abstract Book 61 SESSION VB mOLECULAR THERAPY AND GENOMICS SESSION VB mOLECULAR THERAPY AND GENOMICS September 14, 2012 10:00-11:30 September 14, 2012 10:00-11:30

VB.5: SUBTYPE CHARACTERIZATION OF MALIGNANT PLEURA VB.6: MESOTHELIOMA CELLS METABOLIC DERANGEMENT: A MESOTHELIOMA USING GENE EXPRESSION RATIO TEST NEW MOLECULAR THERAPEUTIC TARGET

Peter E. Sugarbaker1, William G. Richards1, Beow Y. Yeap2, Yaoyu E. Luciano Mutti Wang3, Raphael Bueno1, Assunta De Rienzo1 Onlus, Fondazione Buzzi Per La Ricerca Sul Mesotelioma, Casale 1Division Of Thoracic Surgery, Brigham And Women’s Hospital And Harvard Monferrato (al)/ITALY Medical School, Boston/MA/UNITED STATES OF AMERICA, 2Department Of Medicine, Massachusetts General Hospital And Harvard Medical Introduction: In order to improve the prognosis of patients with Malignant School, Boston/MA/UNITED STATES OF AMERICA, 3Center For Cancer Pleural Mesothelioma (MPM) no question additional many efforts shouid Computational Biology, Dana-Farber Cancer Institute, Boston/MA/UNITED be done to understand if some specific metabolic abnormalities of MPM STATES OF AMERICA do exist and, if they do, how these can exploited for therapeutic purposes. Fondazione Buzzi’s program encompasses a broad range of research september 14, 2012 Background: Malignant Pleural Mesothelioma (MPM) is an uncommon project aimed at bridging the gap between pre-clinical and clinical sciences tumor that can be challenging to diagnose. We previously described in order to set up innovative treatment for this orphan neoplasm.In the utility of the gene ratio technique in discriminating MPM from other particular two projects of this program focused on translation mechanism thoracic malignancies and predicting patient outcome. Herein, we describe and oxidative pattern of mesothelioma cells respectively. for the first time that the gene ratio technique is also able to differentiate the epithelioid from the sarcomatoid type of MPM, a distinction that is Where we are and where we are heading: Translation abnormalities of clinically important for staging and prognosis. In addition, we provide MPM cells: translational implications Previous data suggested that adhesion molecular insights via differential gene expression into pathways and spreading of mesothelioma cells on ECM require the translation of pre- differentially expressed between the epitheliod and the sarcomatoid MPM synthesized mRNAs, and mTORC1 activity and spreading of mesothelioma for further investigation. cells is rapamycin-sensitive and requires continuing translation. More recently it has been showns that the sensitivity of global translation to Methods: We performed gene expression analysis on 39 MPM solid tumors mTOR Inhibition in these same cells depends on the equilibrium between (24 epitheliod, 7 biphasic, and 8 sarcomatoid) using Illumina whole eIF4E and 4E-BP1 but (rather surprisingly) it is not dependent on external genome microarrays. A training set of 8 epithelioid and 8 sarcomatoid growth factors. Moreover many MPM cells are low sensitive to mTOR samples was used to find genes differentially expressed between the two pathway ihibition whereas the sensitivity to mTOR inhibition is driven by groups of samples. Real-Time PCR (RT-PCR) was used to determine the eIF4E and 4E-BP proteins espression. More in details low eIF4E and high relative expression levels of 4 genes (CLDN15, LOC57228, ORF1-FL49, NP), 4E-BP expression are related to mTOR inhibitors sensitivity respectively. and the geometric mean of 3 individual gene pair expression ratios was Hence we evaluated these proteins in specimens from patients with MPM calculated (CLDN15/LOC57228; ORF1-FL49/NP; ORF1-FL49/LOC57228). to predict their sensitivity to MTOR inhibitors.Serine235 phosphorilation The samples were assigned to the epithelioid histology when the combined of translation factor eIF6 plays a key role in murine tumorigenesis and can score was >1 and to sarcomatoid histology when the combined score was be an attractive therapeutic target. IHC staining in specimes from patienst <1. Next, an independent test set of 100 MPMs (63 Epithelioid, 27 Biphasic, with MPM revealed eIF6 overepression in a high number of cases.On the 10 Sarcomatoid) was used for the validation of the test. Functional light of these results eIF6 inhibition either by lentiviral vectors or by PKC enrichment analysis on both Gene Ontology and KEGG was performed beta Kinase inhibithor Enzastaurin has been assessed on in vitro and in using the DAVID web server to identify pathway differentially expressed vivo experiments with promising results that could lead to tailored clinical between the epitheliod and sarcomatoid subtype. treatment of patients with eIF6 overespression Oxidative Energy metabolism of MPM cells: Estrogen Receptor (ERb) is a new therapeutic target for MPM Results: Using the training set expression data, we developed a 4-gene an the other ERb overespressing tumours Female Gender is a well known 3-ratio test able to distinguish the epithelioid from the sarcomatoid MPM positive prognostic factor for patients with MPM and ERb overespression international mesothelioma interest group samples. The test was then validated by RT-PCR in the same 39 MPM has been demonstrated to be associated with better prognosis too. More training set. All the epithelioid and sarcomatoid MPM were correctly recently we also demonstrated how ERb interferes with EFGR espression classified. The same test was then applied using RT-PCR to an independent and affects the sensitivity of MPM cells to EGFR inhibitors. Now we clarified test set of 100 MPM samples showing that 8 of 9 sarcomatoid samples a novel mechanism by which ERb exerts a tumour suppressive effect on (89%) and 62 of 63 (98%) epithelioid MPMs were correctly classified. MPM cells disclosing a novel therapeutic approach to this neoplasm. Both One sarcomatoid sample was excluded from the analysis because the on in vitro and vivo experiments, ERb activation via either overespression result of this test was non diagnostic (1.0). The biphasic MPMs were or treatment with selective agonists significantly compromises distributed in both MPM groups most likely according to relative content of mitochondrial oxidative pathway (affecting mitchondrial sub-units activity) epithelial versus sarcomatoid cells. To identify novel molecular pathways ATP production and decreases cell proliferation and MPM growth.Moreover, specific for the MPM subtypes, we searched for differentially expressed due to the impaired mitochondrial activity, ERb overespressing MPM genes for epitheliod vs. sarcomatoid. We found 183 significant probes cells turn into a much higher dependency on glicolysis and sensitivity to corresponding to 172 genes differentially expressed between the two glicolysis inhibitors. Some selective ERb agonists have been progressively types. The up-regulated biological functions of the 183 probes were related designed over the last few years and are now ready for clinical settings. to transmembrane receptor protein tyrosine kinase signaling, germ cell As ERb espression is common to many human tumours (included prostate development, and regulation of cell proliferation. The down-regulated and breast cancer) the treatment of ERb positive human with selective ERb pathways in the epithelioid group were related to response to external agonists is a novel intriguing approach for a subset of cancer patients stimulus, blood vessel development, cell adhesion, and regulation of secretion. Conclusions: Translation patten and oxidative metabolism of MPM cells seems to disclose a new scenario for the treatment of this tumour even Conclusion: In this study, we show that the gene ratio technique is able to with some implications for “less rare” human tumours.In particular, these distinguish between histological subtypes of MPM with very high sensitivity studies allowed to disclose novel therapeutic biomarkers of sensitivity to supporting the hypothesis that the gene ratio technique may be useful for screen our patients. The next due step is now designing Phase I/II clinical other applications in cancer. trials for patients with MPM selected on the basis detection of biomarkers that could reasonably predict their sensitivity Disclosure: No significant relationships. Disclosure: No significant relationships.

iMig2012.org • Abstract Book 62 Session VC Radiology, Staging and MPM September 14, 2012 10:00-11:30

SESSION VC RADIOLOGY, STAGING AND MPM SESSION VC RADIOLOGY, STAGING AND MPM September 14, 2012 10:00-11:30 September 14, 2012 10:00-11:30 september 14, 2012 VC.2: CT-BASED CLINICAL STAGING FOR MALIGNANT PLEURAL VC.3: IMPACT OF INCORPORATING HEMOGLOBIN LEVEL ON THE MESOTHELIOMA IS UNRELIABLE FOR THE ASSESSMENT OF ACCURACY OF CLINICAL LYMPH NODE STAGING IN MALIGNANT LOCAL TUMOR INVASION AND LYMPH NODE INVOLVEMENT PLEURAL MESOTHELIOMA (MPM).

Thomas Frauenfelder1, Peter Kestenholz2, Roger Hunziker1, Martina William G. Richards1, Ritu R. Gill2, Elizabeth H. Baldini3, Beow Y. Yeap4, Friess2, Walter Weder2, Isabelle Opitz2 Andrea S. Wolf1, Raphael Bueno5, David J. Sugarbaker5 1Division Of Radiology, University Hospital Zurich, Zurich/ 1Division Of Thoracic Surgery, Brigham And Women’s Hospital SWITZERLAND, 2Division Of Thoracic Surgery, University Hospital Zurich, And Harvard Medical School, Boston/MA/UNITED STATES OF Zurich/SWITZERLAND AMERICA, 2Radiology, Brigham And Women’s Hospital, Boston/UNITED STATES OF AMERICA, 3Radiation Oncology, Brigham And Women’s Background: Today computed tomography (CT) remains the most Hospital/Dana-Farber Cancer Institute, Boston/MA/UNITED STATES commonly used imaging modality for malignant pleural mesothelioma OF AMERICA, 4Department Of Medicine, Massachusetts General (MPM) staging, although the visualization of tumor extent remains Hospital And Harvard Medical School, Boston/MA/UNITED STATES OF challenging. The purpose of this study was to assess the reliability of AMERICA, 5Division Of Thoracic Surgery, Brigham And Women’s Hospital, clinical staging for MPM based on preoperative CT scan as a function of Boston/MA/UNITED STATES OF AMERICA different observer and compared to definitive pathological staging. Background: Extrapleural (N2) nodal involvement is an important factor Methods: Sixty two patients with proven MPM and preoperative available that influences the treatment approach for patients with MPM. For patients CT scans were included. Thirty-four patients (55%) underwent talc with involved extrapleural nodes, initial treatment is typically neoadjuvant pleurodesis and all underwent induction chemotherapy prior to surgery. chemotherapy rather than surgery. However, for patients that are node The CT scans were performed over a median of 16 days (0-28) before negative by mediastinoscopy, clinical staging by radiographic imaging surgery using intravenous contrast and had a maximum slice thickness of has low accuracy to detect occult extrapleural nodal disease. We have 2 mm. Three experienced observers blinded to any clinical information identified preoperative anemia as a stage-independent predictor of poor (two chest radiologists and one thoracic surgeon) independently assessed outcome in MPM (Gill, Am J Roentgenol, 2012). We investigated whether clinical T and N stages. Inter-rater reliability was assessed by using analysis preoperative anemia is correlated with pathologic nodal involvement, and of variance. The clinical staging was correlated with definitive pathological might therefore contribute to clinical nodal staging. staging. A kappa statistic was used to assess the agreement of tumor staging between observers. Inter-rater reliability was considered poor (κ ≤ Method: We reviewed a cohort of 168 patients who underwent

0.2), fair (κ = 0.21–0.4), moderate (κ = 0.41–0.6), good (κ = 0.61–0.80) or extrapleural pneumonectomy (EPP) without pre-operative chemotherapy international mesothelioma interest group excellent (κ = 0.81–1.0). at Brigham and Women’s Hospital between 2001 and 2010. Presence of preoperative anemia was determined based on WHO criteria. All Results: The T stage was estimated correctly in 48-71% with a good inter- EPP specimens underwent complete pathologic examination and this observer reliability (κ = 0.64-0.71). The T-stage was underestimated in 22, served as the gold standard for the presence of extrapleural lymph 25 and 37%, respectively, whereas overestimation was rather low (7-15%). node involvement. Nodal status was assessed using the American Joint Regarding the N-status, a correct staging was performed in 58-68%, again Commission on Cancer (AJCC)/International Union Against Cancer (UICC) with good inter-observer agreement (κ = 0.61, 0.65, 0.71). Lymph node TNM classification. Preoperative CT evidence of extrapleural nodal staging was also rather under categorized (16-27%) by all readers. A more involvement was documented by a dedicated thoracic radiologist using size advanced N status was only assigned in 13-16% of the cases. criteria. Frequency of N2 disease in subgroups of patients with and without anemia was compared using Fisher’s exact test. Accuracy of clinical N2 Conclusion: The present set of data confirm that CT scan based clinical staging based on CT was compared to clinical N2 classification plus anemia staging results in an underestimation of the definitive mesothelioma by comparing the relative frequency of correct classification (positive and stage, especially the T stage, which is the most important factor for the negative) by each using a 2-sided McNemar’s test. assessment of resectability of the tumor. Therefore, more precise imaging techniques should be evaluated in comparative studies for a better Results: Median age was 62 (37-79) and 138 patients (81%) were male. prediction of mesothelioma stage of disease. Mediastinoscopy was performed in 133 patients (79%) and was negative for all cases. Histologic subtype of the final specimen was epithelial for 103 Disclosure: No significant relationships. (61%) and non-epithelial for 65 (39%). Extrapleural lymph node disease was identified on final pathologic analysis of the EPP specimen in 53 cases (31%) of which 35 (20%) were associated with preoperative anemia and 18 (11%) were not (p = 0.003). Clinical N2 classification alone was associated with 49% accuracy in predicting pathologic N2 disease (sensitivity 61%, specificity 43%). Clinical N2 classification plus anemia was associated with significantly higher accuracy at 70% (p = 0.003; sensitivity 57%, specificity 76%).

Conclusion: Limited sensitivity of mediastinoscopy for detecting extrapleural N2 nodal disease heightens the urgency to improve current clinical staging strategies for MPM. Preoperative anemia was found to be present in two-thirds of patients with occult N2 disease discovered at final

iMig2012.org • Abstract Book 63 pathology, and has the potential to significantly increase the accuracy of smoking history, chest pain, WBC, hemoglobin, and platelets, univariate radiographic determination of occult N2 disease. analysis revealed smoking, asbestos exposure, weight loss, chest pain and blood values to be significant; however, the stepwise model with Disclosure: No significant relationships. backwards selection for this scenario included only histology, sex, age, WBC, and platelets in the 906 patients with all variables considered.

Conclusion: This is the largest international database examining available SESSION VC RADIOLOGY, STAGING AND MPM prognostic factors which could influence outcomes in surgically managed September 14, 2012 10:00-11:30 MPM patients. Refinement of these models could have clinical utility in defining not only the appropriate patient preoperatively for best outcomes VC.4: SUPPLEMENTARY PROGNOSTIC VARIABLES FOR PLEURAL after cytoreductive surgery, but also in stratifying surgically treated MESOTHELIOMA: A REPORT FROM THE IASLC STAGING patients after clinical and pathologic staging who do or do not receive COMMITTEE adjuvant therapy.

Harvey Pass1, Dori Giroux2, Catherine Kennedy3, Enrico Ruffini4, Disclosure: No significant relationships. september 14, 2012 Ayten K. Cangir5, David Rice6, Hisao Asamura7, David A. Waller8, John Edwards9, Walter Weder10, Hans Hoffman11, Jan P. Van Meerbeeck12, Valerie Rusch13 1Cardiothoracic Surgery, Nyu Langone Medical Center, New York/ SESSION VC RADIOLOGY, STAGING AND MPM NY/UNITED STATES OF AMERICA, 2Crab, WA/UNITED STATES OF September 14, 2012 10:00-11:30 AMERICA, 3University Of Sydney, /AUSTRALIA, 4Dept.Clinical Physiopathology, University Of Torino/ITALY, 5Faculty Of Medicine, Ankara VC.5: DYNAMIC CT AND TUMOR RESPONSE FOR PATIENTS WITH University/TURKEY, 6Thoracic Surgery, MD Anderson Cancer Center, MESOTHELIOMA TX/UNITED STATES OF AMERICA, 7National Cancer Center Hospital/ JAPAN, 8Glenfield Hospital/UNITED KINGDOM, 9University Hospitals Zacariah E. Labby1, William F. Sensakovic1, H.L. Kindler2, Jennifer Sheffield/UNITED KINGDOM, 10Division Of Thoracic Surgery, University Shouldis2, Christopher Straus1, Samuel G. Armato1 Hospital Zurich, Zurich/SWITZERLAND, 11Thoraxklinic/GERMANY, 12Thoracic 1Department Of Radiology, The University Of Chicago, Chicago/IL/UNITED Oncology, Ghent University Hospital, Gent/BELGIUM, 13Department Of STATES OF AMERICA, 2Department Of Medicine, The University Of Chicago, Thoracic Surgery, Memorial Sloan-Kettering Cancer Center, New York/NY/ Chicago/IL/UNITED STATES OF AMERICA UNITED STATES OF AMERICA Background: Dynamic contrast-enhanced (DCE) CT is an imaging modality Background: Current staging for malignant pleural mesothelioma (MPM) that combines the structural information of standard CT with functional is controversial. To plan revisions of this system, the IASLC Staging hemodynamic information. A prospective pilot study is underway to assess Committee developed an international database. Initial analyses focused the viability of DCE-CT as an imaging modality for patients with malignant on surgically-managed patients, and determined that stage, histology, pleural mesothelioma and to investigate correlations between changes sex, age <50 vs. older, and type of procedure (palliative vs. curative) were in DCE-CT metrics and tumor burden measurements in patients on core prognostic variables (IASLC, 2011). The present analysis explores the observation and on chemotherapy. impact of potential, supplementary prognostic variables including the use of adjuvant therapy (i.e., therapy by other modalities including chemotherapy Methods: Patients were scanned with an IRB-approved protocol using a and/or radiation therapy), smoking history, history of asbestos exposure, 256-slice CT scanner. The imaging protocol was designed to efficiently history of weight loss, ECOG performance status, presence of chest utilize one injection of iodinated contrast for both the DCE and full-thorax pain, presence of dyspnea, and preoperative laboratory values including components of the CT scan. The DCE-CT scan consisted of imaging hemoglobin, white blood cell, and platelet counts. snapshots acquired for a 5.5cm axial extent at 120kVp and 100mAs every three seconds for one minute prior to the full-thorax diagnostic scan, international mesothelioma interest group Methods: Participation was solicited from centers with MPM registries. which was followed by five additional snapshots spaced five seconds Common data elements were analyzed by Cancer Research and apart. DCE-CT scans were acquired at each of two sessions approximately Biostatistics (CRAB, Seattle, WA). We analyzed supplementary prognostic 6 weeks apart, at the time of regularly scheduled disease reassessment. variables along with core variables for three scenarios: (A) All variable data All snapshots from a single scan were co-registered using a deformable available, i.e. patient best staged, all core variables, and supplementary registration algorithm to remove the effects of motion prior to the variables are considered (B) Only the clinical stage and the rest of the CORE calculation of DCE-CT parameters. Perfusion, blood volume, time-to-peak, variables excluding surgical staging, and supplementary variables are peak enhancement, and mean transit time data maps were calculated considered, and (C) Supplementary variables excluding adjuvant therapy, using the slope method. Data map spatial statistics were calculated over age, sex, histology, and laboratory parameters are known, i.e., the patient manually defined regions of interest (ROIs) within the tumor, and changes with a diagnosis but no staging performed. Survival was analyzed by Kaplan in data map values between the two DCE-CT scans for each patient were Meier, prognostic factors by logrank and Cox regression model. p<.05 was correlated with tumor burden changes measured according to the Modified significant. RECIST technique as well as volumetric disease measurements.

Results: Data included 3101 patients (15 centers, 4 continents) from 1/95- Results: Thirteen mesothelioma patients (seven on observation, six on 8/09 of which 2141with best (clinical or pathologic staging recorded or chemotherapy) have undergone two DCE-CT scans to date in this ongoing both) TNM stages had non-missing age, sex, histology, and type of surgical study. Tumor ROIs exhibited heterogeneous contrast uptake, ranging from procedure. When all variable data were considered (Scenario A), univariate no uptake to peak enhancement of 100HU. No DCE-CT parameters were analysis revealed adjuvant therapy, asbestos exposure, weight loss, PS, significantly correlated with changes in tumor burden in this initial pilot chest pain, hemoglobin (<14.6 or not), platelet count (> 400k or not, and WBC(> 15.5 or not) as significant supplementary variables. Stepwise Cox Regression Model after elimination of non-significant variables revealed best stage, histology, sex, age, type of surgery, adjuvant treatment, WBC, and platelets in a final model (n=555 patients with all variables under consideration.) When Clinical Stage was considered with other CORE and supplementary variables (Scenario B), stepwise Cox Regression Model with backwards selection included clinical stage, histology, sex, age, type of surgery, adjuvant treatment, WBC, hemoglobin, and platelets in 627 patients. When limited data are considered at patient presentation (Scenario 3) including age, sex, histology, asbestos exposure, weight loss,

iMig2012.org • Abstract Book 64 shrinking lung sign (sensitivity 50%, specificity 85.7% and accuracy 63.2%); the positive predictive value was 85.7%.The sign of pointillism, however, could diagnose 68 patients correctly (sensitivity 91.7%, specificity 85.7% and accuracy 89.5%), with a positive predictive value of 91.7% and negative predictive value of 85.7%. september 14, 2012

cohort. However, there was a marked difference in DCE-CT temporal Conclusion: This new sign of pointillism seems to be a good predictor perfusion changes between the observation cohort (n=7) and the treatment for malignant pleural disease; especially for MPM it can provide guidance cohort (n=6) (+29% and -13%, respectively). for thoracoscopic evaluation and possibly avoiding unnecessary invasive procedures. As expected, the shrinking lung sign has a good positive Conclusion: This study indicates that DCE-CT provides functional predictive value. hemodynamic information for mesothelioma and that changes in calculated DCE-CT parameters may provide imaging biomarkers of underlying tumor Disclosure: No significant relationships. change not reflected in tumor measurements. Accrual continues until 20 patients (10 on observation, 10 on chemotherapy) have each received two DCE-CT scans. SESSION VC RADIOLOGY, STAGING AND MPM Disclosure: No significant relationships. September 14, 2012 10:00-11:30

VC.7: THE IMPACT OF PET SCANNING IN THE MANAGEMENT OF PATIENTS WITH LOCALLY ADVANCED MALIGNANT PLEURAL SESSION VC RADIOLOGY, STAGING AND MPM MESOTHELIOMA RECEIVING HIGH-DOSE HEMITHORACIC September 14, 2012 10:00-11:30 RADIOTHERAPY VC.6: THE SIGN OF POINTILLISM ON DWI INCREASES THE Malcolm Feigen1, Sze Ting Lee2, Robert Fabiny3, Andrew M. Scott2, ACCURACY OF MPM DIAGNOSIS Chris Hamilton1 1Radiation Oncology Center, Austin Health, Heidelberg West/VIC/ Johan Coolen1, Frederik De Keyzer1, Philippe Nafteux2, Walter De AUSTRALIA, 2Centre For Pet, Ludwig Institute For Cancer Research, Austin Wever1, Christophe Dooms3, Eric Verbeken4, Paul De Leyn2, Dirk Van Health, /VIC/AUSTRALIA, 3Radiology Dept, Austin Health/AUSTRALIA Raemdonck5, Johan Vansteenkiste3, Kristiaan Nackaerts6, Steven Dymarkowski7, Johny Verschakelen1 1 2 Background: Imprecise imaging of all active tumor sites is a fundamental

Radiology, Uz Gasthuisberg, Leuven/BELGIUM, Thoracic Surgery, international mesothelioma interest group 3 problem in the management of malignant pleural mesothelioma, where CT University Hospital, Leuven/BELGIUM, Pneumology, Az Gasthuisberg, scans have poor specificity and misinterpret regions of benign fibrosis. We Leuven/BELGIUM, 4Anatomopathology, Az Gasthuisberg, Leuven/ 5 analyzed patients with locally advanced mesothelioma who were registered BELGIUM, Thoracic Surgery, Az Gasthuisberg, Leuven/BELGIUM, 18 6 7 on our FDG-PET/CT scan database and received high-dose radiotherapy Pneumonology, Az Gasthuisberg, Leuven/BELGIUM, Radiology, Az (RT), to assess how the scans contributed to patient outcomes. Gasthuisberg, Leuven/BELGIUM Methods: In 2003 our institution commenced a program of high-dose Background: The diagnosis of malignant pleural mesothelioma (MPM) RT for selected patients with localized pleural mesothelioma, requiring should be considered in any patient with either pleural fluid or pleural potential patients to have a staging PET scan prior to RT simulation. We thickening, especially if chest pain is present. However, pathological aimed to obtain a followup PET scan 3 months post-RT, and whenever confirmation of MPM is not always straightforward, as both on clinical further intervention was considered. PET scans were co-registered with examination and histopathology MPM can present as a wolf in sheep’s simulation CT scans and the planning target volume (PTV) was compared clothing. Therefore, we prospectively evaluated the potential role of on all subsequent PET scans to measure total glycolytic volumes and assess magnetic resonance imaging in this setting. For this study purpose we any 18FDG-avidity at other sites. visually assessed two radiological parameters, namely the ‘shrinking lung’ sign and the newly introduced sign of pointillism on diffusion-weighted Results: Between July 2003 and March 2012, 27 patients received RT imaging (DWI). to a dose of 45-60 Gy to part or all or the hemithorax and had 82 PET scans available for analysis, from 3 to 89 (median 8) months post-RT. Methods: Seventy-six consecutive patients with pleural abnormalities Most patients were males (81%) with right-sided (59%) epithelioid (78%) suspicious for MPM underwent MRI, including DWI and dynamic contrast- mesotheliomas. Before RT, 16 underwent pleurectomy/decortication, 8 enhanced MRI (DCE-MRI), additional to the clinical imaging, explorative had a pleurodesis and 2 biopsy only. 15 received chemotherapy, including thoracoscopy and histopathological confirmation. Both CT and MR data 4 planned for trimodality therapy. One patient who had an extrapleural were correlated with pathology. Because most clinicans believe in the high pneumonectomy recurred after adjuvant chemotherapy prior to RT. 83% diagnostic value of visual parameter interpretation we correlated the sign had clinical stage III/IV disease, as determined by their initial PET scan. 11 of shrinking lung (i.e. volume decrease of hemithorax due to fibrosis) with received 3D-conformal RT and 16 intensity-modulated RT. Eight patients the sign of pointillism (i.e. hyperintensities on DWI images acquired with a 2 are living, 4 disease-free at median followup of 15.3 months. The planning high b-value of 1000 s/mm ). PET scan identified new sites of disease beyond the hemithorax in 8% of patients, who were withdrawn from our program as they were considered Results: This cohort existed of 28 patients with benign pleural alterations at high risk of developing distant disease, and 58% of those selected had and 48 patients with malignant pleural diseases, of which 42 MPM no new metastases within 6 months of RT. Areas of high FDG uptake were (55.3%). A total of 48 patients were diagnosed correctly by using the

iMig2012.org • Abstract Book 65 planned to receive concurrent radiation boosts to higher doses, resulting in improved local control rates. Patterns of failure based on PET and CT scan analyses confirmed a high rate of locoregional control of mesothelioma after RT, with only 4 patients relapsing in-field, all but one with concurrent metastases documented in unirradiated sites. Five patients remain disease-free and 18 developed out-of-field recurrences. One patient with an a PET-detected solitary recurrence in ipsilateral hilar nodes outside the volume irradiated 5.5 years earlier remains in remission 3 years after salvage RT and 8 years after diagnosis. Changes in patterns of radiation pneumonitis over time will be addressed.

Conclusion: In our mesothelioma program, PET/CT scans play a pivotal role in patient selection for surgery and high-dose RT, radiotherapy planning that targets sites of gross unresected disease with higher doses and treatment followup to detect recurrences for early salvage therapies. Based september 14, 2012 on followup PET scans confirming recurrent disease within the radiation target volume in only 4 of 27 patients, we can confirm that radiotherapy produces effective locoregional control in selected mesothelioma patients who receive high-dose irradiation, a locoregional control rate of 85%.

Disclosure: No significant relationships. international mesothelioma interest group

iMig2012.org • Abstract Book 66 Workshop XI miRNA and Mesothelioma September 14, 2012 13:15-14:15

WORKSHOP XI mIRNA AND MESOTHELIOMA WORKSHOP XI mIRNA AND MESOTHELIOMA September 14, 2012 13:15-14:15 September 14, 2012 13:15-14:15 september 14, 2012 WSXI.1: SPECIMEN SELECTION FOR MICRORNA QUANTIFICATION WSXI.2: MICRORNAS AS NOVEL DIAGNOSTIC BIOMARKERS IN MESOTHELIOMA AND REGULATORS OF TUMOR BIOLOGY IN MALIGNANT MESOTHELIOMA Jenette Creaney1, Ian Dick1, Justine Leon1, Yvonne Demelker1, Arthur W. Musk2, Bruce W.S. Robinson3 Bahareh Badrian1, Kimberly A. Birnie2, Y C G. Lee2, Glen Reid3, Nico Van 1School Of Medicine And Pharmacology, University Of Western Australia, Zandwijk3, Bruce W.S. Robinson4, Arthur W. Musk5, Jenette Creaney6, Nedlands/AUSTRALIA, 2Respiratory Medicine, Sir Charles Gairdner Hospital, Steven E. Mutsaers7 Nedlands/WA/AUSTRALIA, 3School Of Medicine And Pharmacology, 1Lung Institute Of Western Australia, Perth/AUSTRALIA, 2Lung Institute Of University Of Western Australia, Perth/WA/AUSTRALIA W.A. University Of Western Australia, Perth/WA/AUSTRALIA, 3Asbestos Diseases Research Institute, University Of Sydney, Sydney/ Background: There is a lot of interest in the use of microRNA (miRNA) AUSTRALIA, 4School Of Medicine And Pharmacology, University Of profiles as diagnostic tools in mesothelioma and several candidate miRNA Western Australia, Perth/WA/AUSTRALIA, 5Respiratory Medicine, Sir biomarkers have been described. Recently we reported that circulating Charles Gairdner Hospital, Perth/AUSTRALIA, 6School Of Medicine And levels of the mir-625-3p were significantly elevated in the plasma and Pharmacology, University Of Western Australia, Perth/AUSTRALIA, 7Lung serum of mesothelioma patients relative to controls. Despite the intrinsic Institute Of Western Australia, Perth/WA/AUSTRALIA stability of miRNAs there are concerns relating to the variability of miRNA quantification because of other factors including specimen collection Background: There is currently no internationally uniformly accepted methods, RNA extraction efficiency and data analysis. In the present study approach for the diagnosis of Malignant Mesothelioma (MM). The success we compared miRNA levels in longitudinal blood samples collected by rate of current methods for diagnosis is highly variable and at some centers two different methods and also examined the diagnostic accuracy of our definitive diagnosis can take up to 3 months. Therefore, there is an urgent candidate miRNA biomarker mir-625-3p compared to soluble mesothelin need for identification of new biomarkers in MM. MicroRNAs (miRs) are levels. single stranded RNAs which regulate gene expression and have been shown to be important in cancer pathogenesis. miRs can be easily measured, Methods: Blood samples were initially collected from healthy controls are expressed in body fluids such as serum and sputum and are extremely on three consecutive days. Parallel samples were collected in traditional stable. Therefore, miRs represent an attractive target for biomarker “red cap” serum BD Vacutainers and in PAXGene Blood RNA Tubes discovery in MM.Therefore the aim of this study is to profile serum and cells (Qiagen). Serum collected in traditional tubes were allowed to clot at isolated from Pleural Effusions (PE Cells) from MM patients for all identified room temperature before being stored at -20oC and miRNA extracted on human miRs and select potential biomarker targets.

the batch of samples using the protocol of Miska et al. 2002. Samples international mesothelioma interest group collected in PAXgene tubes were processed following the manufacturer’s Methods: Total RNA was isolated from 50 PE Cells (30 MM, 10 Benign and instructions. MiRNA was quantitiated using TaqMan® MicroRNA Assays on 10 Adenocarcinoma (AC)) and 60 serum (30MM, 10 asbestos exposed but the Applied Biosystems StepOne Plus Real Time PCR System. Blood samples healthy, 10 AC and 10 asbestosis) samples.The OpenArray® Real-Time were prospectively collected in PAXGene Blood RNA Tubes from 25 patients PCR platform was used to profile the samples for miRs. Data analysis was with malignant mesothelioma and 25 patients with asbestos related benign performed in DataAssist. For validation of selected targets, miRs were disease. MiRNA was extracted and quantitated as above. measured using qPCR.

Results: Levels of the small nuclear RNAs U44 and U6B, and the mirs-16, Results: The miR targets were selected based on expression profile and -103, -192 and -21 showed significantly greater variance in a mixed model significance of expression when MM was compared to the other control component analysis in blood samples collected in serum tubes from four groups. Five novel miR targets were selected from serum and 5 from PE health individuals on three consecutive days compared to parallel samples Cells. These 10 targets were initially validated by qPCR in the same samples collected in PAXGene tubes. Levels of mir-625-3p were significantly to confirm the profiling results. To determine the diagnostic potential of elevated in samples collected in PAXGene tubes from mesothelioma these selected targets qPCR is currently being performed in a larger cohort patients relative to controls. Levels of soluble mesothelin were significantly of 100 samples. Furthermore, using our data we were able to validate miRs elevated in the serum of mesothelioma patients relative to controls. There identified by other studies which also strengthens our profiling data. was no significant difference between the diagnostic accuracy of the two biomarkers when the area under the receiver operator curves was Conclusion: Currently this work is the largest and most comprehensive compared. Data will be presented from additional 30 patients to increase miR profiling study in MM. We have identified a number of novel miR the power of the study. biomarkers in serum and PE samples. The results of this study may not only identify a biomarker which will improve current diagnostic procedures but Conclusion: MiRNA levels are more stable when the miRNA is extracted one that can be used to screen high risk individuals and ultimately improve from serum in PAXgene tubes, as opposed to normal serum tubes. By using patient outcomes. the PAXgene tubes, consistent miR levels over consecutive days can be seen and would therefore prove to be a more reliable source when it comes Disclosure: No significant relationships. to diagnosing patients with malignant mesothelioma.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 67 WORKSHOP XI mIRNA AND MESOTHELIOMA WORKSHOP XI mIRNA AND MESOTHELIOMA September 14, 2012 13:15-14:15 September 14, 2012 13:15-14:15

WSXI.3: ANALYSIS OF MICRO-RNAS AND GENES IN MALIGNANT WSXI.4: IDENTIFICATION OF MIR-625-3P AS POTENTIAL BLOOD- MESOTHELIOMA BASED BIOMARKER FOR MALIGNANT PLEURAL MESOTHELIOMA

Chuong D. Hoang1, Purvesh Khatri1, Yue Xu1, Robert Merritt1, Joseph Michaela B. Kirschner1, Yeun Yee Cheng1, Bahareh Badrian2, Steven C. Shrager1, Heather Wakelee1, Atul Butte1, Robert A. Kratzke2 Kao1, Jenette Creaney3, J.J.B. Edelman4, Nicola J. Armstrong5, Michael P. 1Stanford University, Stanford/CA/UNITED STATES OF AMERICA, 2Medicine, Vallely4, Arthur W. Musk6, Bruce W.S. Robinson7, Brian C. Mccaughan4, Div Of Hematology, Oncology, And Transplantation, University Of Sonja Klebe8, Steven E. Mutsaers2, Nico Van Zandwijk1, Glen Reid1 Minnesota, Minneapolis/MN/UNITED STATES OF AMERICA 1Asbestos Diseases Research Institute, University Of Sydney, Concord/ AUSTRALIA, 2Lung Institute Of Western Australia, Centre For Asthma, Background: We hypothesized that certain microRNA (miRNA)-mRNA Allergy, And Respiratory Research And Pathwest Laboratories Of Medicine, interactions are associated with malignant transformation and progression University Of Western Australia, Nedlands/AUSTRALIA, 3School Of of malignant mesothelioma (MM). To date, no integrated analysis has been Medicine And Pharmacology, University Of Western Australia, Perth/ september 14, 2012 performed to identify critical interactions between miRNA-mRNA that may AUSTRALIA, 4Cardiothoracic Surgical Unit, Royal Prince Alfred Hospital; The drive the MM malignant phenotype. Baird Institute And Faculty Of Medicine, University Of Sydney, Newtown/ AUSTRALIA, 5Garvan Institute For Medical Research And School Of Methods: Total RNA was extracted from 22 specimens of biopsy proven Mathematics And Statistics, University Of New South Wales, Darlinghurst/ human MM and 6 specimens of normal pleura. Paired global transcriptional AUSTRALIA, 6Respiratory Medicine, Sir Charles Gairdner Hospital, profiles of miRNA and genes (mRNA) expression were generated using Nedlands/WA/AUSTRALIA, 7School Of Medicine And Pharmacology, Illumina microarrays. After rank-invariance normalization of miRNA and University Of Western Australia, Perth/WA/AUSTRALIA, 8Department Of mRNA expression data, we used significance analysis of microarray (SAM) Anatomical Pathology, Flinders University, Adelaide/AUSTRALIA to identify differentially expressed miRNAs and mRNAs at a false discovery rate (FDR q-values) < 5%. To better determine relevant genes for a given Background: The definitive diagnosis of malignant pleural mesothelioma process, sets of genes whose expression levels negatively correlated with (MPM) often depends on the availability of a biopsy of sufficient size. specific miRNAs were identified as likely direct targets of that miRNA. The identification of a biomarker that can be easily measured in blood These differentially expressed miRNAs and mRNAs were analyzed using would represent an important step forward. Recently it has been shown Ingenuity Pathway Analysis (IPA) to identify the biologic and chemical that microRNAs (miRNAs) detectable in serum/plasma represent a class systems that are affected by coordinately altered levels of miRNAs of potential new biomarkers. In this study we investigated the ability of and mRNAs in MM. We integrated differentially expressed miRNAs and miRNAs in plasma/serum to serve as a diagnostic marker for MPM. mRNAs at the pathway, systems level using multiple miRNA-mRNA target databases from the public domain. We constructed an interaction network Methods: Using Agilent 8x15k miRNA microarrays we profiled miRNA of differentially expressed miRNA-mRNA pairings. expression in plasma samples from healthy volunteers and patients with MPM. Candidate miRNAs identified in the arrays were validated by TaqMan Results: We found 2,202 genes (593 up, 1609 down) differentially assay-based quantitative real-time PCR or using the OpenArray real-time expressed in MM specimens compared to normal pleura using SAM. PCR platform. Similarly, we identified 101 miRNAs differentially expressed (57 down, 94 up). Focusing on negatively correlated pairings of miRNA-mRNA, Results: Microarray-based expression profiling of plasma from 5 MPM we generated 2 datasets for further analysis. As an example, when patients and 3 healthy controls identified 17 miRNAs with significantly we analyzed the set of down-regulated miRNA in MM as compared to differential abundance in the two sample groups. Validation of these normal pleura using IPA, there was over-representation of these miRNA miRNAs in a series of plasma samples from 15 MPM patients and 13 in immune response-related biologic pathways including immune cell controls (healthy individuals and patients with coronary artery disease) trafficking, cellular motility, antigen presentation, cell-to-cell signaling revealed that levels of miR-625-3p, were significantly elevated in plasma international mesothelioma interest group and interaction, and cellular morphology. Complementary analysis of MPM patients (4-fold, p=0.004), and able to discriminate between MPM of the 593 over-expressed genes by the IPA algorithm showed similar patients and controls (sensitivity of 73.3 %, specificity 78.6 %). Levels of enrichment in immune response-related pathways. We used miRNA-mRNA two miRNAs previously reported to be associated with MPM, miR-29c* target databases from the public domain to build an interaction network and miR-92a, were also elevated in our MPM series however without using under-expressed miRNAs and over-expressed mRNAs. We mapped reaching statistical significance. Assessing levels of miR-625-3p in serum onto these networks confirmed expression data from our group of 28 of an independent series of MPM (N=30) and asbestosis (N=10) patients specimens. We observed a large number of miR-1 gene targets over- confirmed that miR-625-3p was significantly (p=0.023) elevated only in expressed in MM, while miR-1 itself was significantly under-expressed serum of MPM patients and was able to discriminate between cases and (FDR < 0.05). Expression of miR-1 negatively correlated with expression of controls with a sensitivity of 70 % and a specificity of 90 %. Finally, miR- 52 mRNA (FDR < 0.05). This miRNA was underexpressed in MM by 3-fold 625-3p was present at significantly elevated levels (2-fold, p=0.006) in more (FDR < 0.01), while its gene targets showed an opposite pattern being tumour specimen from 18 MPM patients compared to normal mesothelium overexpressed in MM. In addition to positively affecting inflammation- (pericardial tissue). Preliminary data from a third series of serum samples related pathways, miR-1-related gene subsets were enriched for oncogenic from 32 MPM patients and matched healthy controls so far confirm our genes (e.g. p21, TRAF2, and SAA1) positively impacting cell growth, observation that miR-625-3p is elevated at least 3-fold in blood from survival, and anti-apoptosis. MPM patients. Final results from this third series will be presented at the conference. Conclusion: Integrated analysis of miRNA and mRNA expression profiles in MM revealed novel miRNA associated with MM and identified putative Conclusion: Taken together the data from the initial two series of blood interactions that may underlie the malignant phenotype. These miRNA- samples provide evidence that miR-625-3p has the potential to serve as mRNA are candidates for functional validation. miR-1 is a putative tumor a novel blood-based biomarker for MPM. Preliminary results from a third suppressor miRNA in MM. series of serum samples support these findings.

Disclosure: No significant relationships. Disclosure: No significant relationships.

iMig2012.org • Abstract Book 68 WORKSHOP XI mIRNA AND MESOTHELIOMA September 14, 2012 13:15-14:15

WSXI.5: THE MICRORNA MIR-16 IS A NOVEL TUMOUR SUPPRESSOR GENE IN MALIGNANT PLEURAL MESOTHELIOMA

Glen Reid1, Marcella Pel2, Michaela B. Kirschner1, Marissa Williams1, Casey M. Wright1, Yeun Yee Cheng1, Brian C. Mccaughan3, J.J.B. Edelman3, Michael P. Vallely3, Rayleen V. Bowman4, Sonja Klebe5, Nico Van Zandwijk1 1Asbestos Diseases Research Institute, University Of Sydney, Concord/AUSTRALIA, 2Academic Medical Centre, Amsterdam/ NETHERLANDS, 3Cardiothoracic Surgical Unit, Royal Prince Alfred Hospital; The Baird Institute And Faculty Of Medicine, University Of Sydney, Newtown/AUSTRALIA, 4Department Of Thoracic Medicine, The Prince september 14, 2012 Charles Hospital, Brisbane/QLD/AUSTRALIA, 5Department Of Anatomical Pathology, Filnders Univerity, Adelaide/AUSTRALIA

Background: The microRNA expression in malignant pleural mesothelioma (MPM) biopsy specimens and cell lines is significantly altered. Previous studies have indicated that a number of microRNAs have tumour- suppressor or oncogenic functions in MPM. The expression of miR-16 is frequently lost in a variety of cancers, and here we demonstrate that miR-16 is also down-regulated in MPM and seems to function as a tumour- suppressor gene.

Methods: Tumour content of formalin-fixed, paraffin-embedded specimens from patients who underwent extrapleural pneumonectomy was marked by an experienced pathologist prior to laser-capture micro- dissection and RNA isolation. The expression of selected microRNAs was measured and compared with levels in formalin-fixed pericardial tissue using miR-specific TaqMan assays. Following RNA extraction with TRIzol, the expression of miR-16 was also measured in MeT-5A and a panel of MPM cell lines. Re-expression of miR-16 was accomplished by transfecting cells with a miR-16 mimic. Effects on growth and gemcitabine resistance were measured using a SYBR Green-based proliferation assay, and colony formation assays were also carried out. The expression of potential miR-16 targets in MPM was analysed by RT-qPCR in miR-16 mimic-transfected cells.

Results: The expression of miR-16 was significantly (p = 0.01) down- regulated (20-fold) in MPM tumours compared with pericardial tissue. In cell lines, miR-16 expression in MPM lines was 2 to 4-fold down-regulated compared with MeT-5A, an immortalized normal mesothelial cell line. international mesothelioma interest group Transfection with a miR-16 mimic increased intracellular levels of mature miR-16 in all transfected lines. The restoration of miR-16 expression resulted in growth inhibition in all MPM lines in a dose-dependent manner, with as little as 1 nM mimic leading to significant effects on proliferation and the ability to form colonies when seeded at low density. In contrast, transfection with as much as 5 nM did not effect the growth of MeT-5A cells. The transfection of MPM cells with miR-16 also led to a 2 to 5-fold sensitization to gemcitabine but did not affect MeT-5A sensitivity to this drug. The expression of putative miR-16 targets related to gemcitabine resistance were down-regulated in transfected cells.

Conclusion: The down-regulation of miR-16 in MPM tumour samples and cell lines, together with the growth inhibitory effects of restoring miR- 16 expression suggests a tumour suppressor function of this microRNA in MPM. Together with its ability to sensitize MPM cells to gemcitabine treatment, this suggests miR-16 replacement should be explored as a novel therapeutic approach in MPM.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 69 Session VIA Novel Therapeutics: Preclinical Studies September 14, 2012 14:20-15:50

SESSION VIA NOVEL THERAPEUTICS: PRECLINICAL STUDIES SESSION VIA NOVEL THERAPEUTICS: PRECLINICAL STUDIES September 14, 2012 14:20-15:50 September 14, 2012 14:20-15:50 september 14, 2012 VIA.1: TARGETING THE EXTENDED ERBB RECEPTOR FAMILY VIA.2: ZOLEDRONIC ACID, THE THIRD GENERATION OF USING CHIMERIC ANTIGEN RECEPTOR (CAR)-TARGETED T-CELLS BISPHOSPHONATES, PRODUCES ANTI-TUMOR EFFECTS ON IN MALIGNANT PLEURAL MESOTHELIOMA (MPM) CELL LINES. MESOTHELIOMA IN VITRO AND IN VIVO THROUGH APOPTOSIS OR S PHASE ARREST IN P53-INDEPENDENT AND RAS 1 2 1 Ana C. Parente Pereira , Astero Klabatsa , Leticia Bosshard-Carter , PRENYLATION-INDEPENDENT MANNERS May Van Schalkwyk1, Sjoukje Van Der Stagen1, Thivyan Thayaparan1, 1 John Maher Yuji Tada1, Quanhai Li2, Kiyoko Kawamura2, Hiroshi Kobayashi3, Ikuo 1 Research Oncology, Division Of Cancer Studies, King’s College London, Sekine4, Yuichi Takiguchi4, Koichiro Tatsumi1, Hideaki Shimada5, Kenzo 2 London/UNITED KINGDOM, Thoracic Surgery, Guy’s And St Thomas’ Nhs Hiroshima6, Masatoshi Tagawa7 Foundation Trust, London/UNITED KINGDOM 1Respirology, Chiba University School Of Medicine, Chiba/ JAPAN, 2Pathology And Cell Therapy, Chiba Cancer Center Research Background: MPM is a lethal malignancy caused by asbestos which Institute, Chiba/JAPAN, 3Biochemistry, Graduate School Of Pharmaceutical affects around 2,000 people in the UK annually. It is characterised by Sciences,Chiba University, Chiba/JAPAN, 4Medical Oncology, Chiba poor prognosis and resistance to chemotherapy. Currently, treatment University, Chiba/JAPAN, 5Surgery, Toho University School Of Medicine, options include multimodality therapy with a combination of surgery, Tokyo/JAPAN, 6Pathology, Tokyo Women’s Medical University Yachiyo chemotherapy and radiotherapy, but they fail to give a survival benefit of Medical Center, Chiba/JAPAN, 7Division Of Pathology And Cell Therapy, more than a few months. It is therefore necessary that new therapeutic Chiba Cancer Center Research Institite, Chiba/JAPAN approaches are developed. Here we present a novel immunotherapeutic approach targeting the extended ErbB receptor family using CAR-targeted Background: Bisphosphonates have a strong affinity for mineralized bone T-cells in MPM cell lines. The ErbB family consists of EGFR, HER-2, ErbB-3 matrix and inhibit bone absorption by acting on osteoclasts, and are and ErbB-4, which undergo ligand-induced homo- and heterodimerization. currently used for bone lesions such as osteoporosis and hypercalcemia. Over-expression of EGFR in seen in the majority of MPM and ER-2 is also The third generations of bisphosphonates inhibit farnesyl pyrophosphate commonly expressed. Emerging evidence suggests that ErbB3 might synthetase, a key enzyme in the mevalonate pathways, and deplete also be an important player in MPM. We have previously developed an isoprenoid pools, which subsequently result in decreased prenylation immunotherapeutic approach that targets several ErbB dimers. A chimeric of small G proteins. The unprenylation influences activities of the small antigen receptor (CAR, named T1E28z) was engineered in which the G proteins, which plays a crucial role in a variety of biological functions promiscuous ErbB ligand, T1E, is fused to a CD28+CD3ζ endodomain. As including cell survival. poor transduction efficiency of patient-derived T-cells may be a limiting

factor in achieving clinical efficacy, a chimeric cytokine receptor (4αβ) in Methods: We examined whether zoledronic acid (ZOL), the third international mesothelioma interest group which the ectodomain of IL-4 receptor α was fused to the endodomain of generation of bisphosphonates, produced cytotoxic effects on human the shared β receptor used by IL-2 and IL-15 was constructed. T-cells that mesothelioma cells, and investigated a possible involvement of p53, Ras express 4αβ can be selectively expanded using IL-4, greatly facilitating and extracellurar signal regulated kinase1/2 (ERK1/2) pathways. Cytotoxicity the rapid generation of therapeutic cell products. Burt et al (2012) have and cell cycle were assessed with colorimetric assay and flow cytometry, recently shown that MPM progression is driven in part by IL-4, produced respectively. Expression levels of apoptosis-linked proteins and prenylation by T-cells within the tumour (1). Using the4αβ system described above, of small G proteins were tested with p53-siRNA, an ERK kinase1/2-inhibitor we could capitalise upon local IL-4 to provide targeted support for our and prenyl alcohols. The anti-tumor activity was examined in an orthotopic engineered T-cells, directly at the site of the tumour. animal model.

Methods: Peripheral blood mononuclear cells were isolated from healthy Results: ZOL treatments suppressed growth of mesothelioma cells bearing controls and activated using CD3/CD28 beads. Co-expression of T1E28z the wild-type p53 gene through apoptosis induction accompanied by and 4ab (named “T4”) was achieved using a single SFG retroviral vector activation of caspases-8, -9 and -3, or S-phase arrest by up-regulated and delivered to the T-cells. Cells were expanded over a period of 15 days cyclin A and B1. ZOL elevated p53 expression levels and the p53 in the presence of IL-4 for specific expansion of T4. Co-cultures were set up phosphorylation at Ser 15 residues, which subsequent activation of the to test T4 efficacy in vitro. The mesothelioma cell models used were H28, downstream pathways and decreased mitochondrial membrane potentials. REN and MM98. Conditional media was collected to analyse INF-gamma Down-regulated p53 expression with the siRNA however showed that production in order to test for T-cell activation. both apoptosis and S-phase arrest were irrelevant to the p53 activation, demonstrating that p53 pathways were not involved in the ZOL-mediated Results: Destruction of cell monolayers derived from all three cell lines cytotoxicity. In contrast, geranylgeranyl but not farnesyl pyrophosphate was achieved within 24h upon co-cultivation with T4 transduced T-cells. inhibited ZOL-induced apoptosis and S-phase arrest, suggesting that Tumour destruction and INF-gamma production show specific activation of the cytotoxic activity was attributable to unprenylated small G proteins. the T4 cells upon meeting the target. Moreover, geranylgeraniol supplement decreased ZOL-mediated Rap1A but not Ras unprenylation, and inhibition of ERK1/2 pathways suppressed ZOL- Conclusion: Therapeutic efficacy has been demonstrated in vitro following induced apoptosis but not S-phase arrest. We further demonstrated that delivery of T4 T-cells to monolayer MPM cultures, providing all the ZOL, administered intrapleurally, inhibited the tumor growth developed in necessary data for in vitro work with patients and development of in the pleural cavity without inducing significant adverse effects. vivo models. 1. Burt BM, Bader A, Winter D, Rodig SJ, Bueno R, Sugarbaker DJ. Expression of interleukin-4 receptor alpha in human pleural Conclusion: ZOL induced apoptosis or S-phase arrest, both of which are mesothelioma is associated with poor survival and promotion of tumor independent of p53 activation and Ras unprenylation. ZOL achieved anti- inflammation. Clin Cancer Res. 2012 Mar 15; 18(6):1568-77

Disclosure: No significant relationships. iMig2012.org • Abstract Book 70 tumor effects and is a possible therapeutic agent to mesothelioma when SESSION VIA NOVEL THERAPEUTICS: PRECLINICAL STUDIES administered in the pleural cavity. September 14, 2012 14:20-15:50

Disclosure: No significant relationships. VIA.4: NOVEL INTERNALIZING HUMAN ANTIBODY-TARGETED INTRACELLULAR DELIVERY OF SMALL RNA THERAPEUTICS TO ALL SUBTYPES OF MESOTHELIOMA

SESSION VIA NOVEL THERAPEUTICS: PRECLINICAL STUDIES Yang Su, Scott Bidlingmaier, Bin Liu September 14, 2012 14:20-15:50 UCSF, San Francisco/CA/UNITED STATES OF AMERICA

VIA.3: A MULTIFUNCTIONAL MESOTHELIN ANTIBODY-TAGGED Background: The goal of this study is to develop effective small interfering MICROPARTICLE TARGETS HUMAN MESOTHELIOMAS RNA (siRNA) therapeutics against mesothelioma. As most mammalian cells do not actively take up siRNA, therapeutic application of siRNAs Brooke T. Mossman1, Sherrill L. Macura1, Jedd M. Hillegass1, Jeremy L. requires the development of new delivery methods. The work is built Steinbacher2, Arti Shukla1, Kelly J. Butnor1, Douglas J. Taatjes1, Risto A. on our previous studies where we used phage antibody library selection september 14, 2012 Kauppinen3, Raffit Hassan4, Christopher C. Landry2 approach to identify mesothelioma-targeting antibodies, and discovered 1Pathology, University Of Vermont College Of Medicine, Burlington/ a panel of novel internalizing human single chain antibodies (scFvs) that VT/UNITED STATES OF AMERICA, 2Chemistry, University Of Vermont, target all subtypes of mesothelioma. We hypothesize that our panel of Burlington/VT/UNITED STATES OF AMERICA, 3Geisel School Of mesothelioma-targeting internalizing scFvs are well suited for developing Medicine, Dartmouth Medical School, Hanover/NH/UNITED STATES OF effective vehicles for systemic siRNA delivery. This study aims to achieve AMERICA, 4Laboratory Of Molecular Biology, National Cancer Institute, mesothelioma-targeted intracellular delivery of small RNA therapeutics in Bethesda/MD/UNITED STATES OF AMERICA vitro and in vivo.

Background: Pleural and peritoneal mesotheliomas (MMs) are Methods: We first modified our internalizing scFvs to impart them with chemoresistant tumors with no effective therapeutic strategies. We siRNA binding and delivery function. We identified a series of small RNA hypothesized that local injections of porous microparticles loaded with binding motifs (SRBMs) and genetically fused them with internalizing scFvs. chemotherapeutic drugs and externally modified using an antibody for We constructed expression vectors and identified scFv-SRBMs that can be mesothelin (MB) would be valuable in treating MMs. well expressed and readily purified. For quality control, we tested purified scFv-SRBM fusions for binding to mesothelioma cells by flow cytometry, Methods: We first prepared multifunctional, acid-prepared mesoporous and studied internalization by confocal microscopy. We studied potency spheres (APMS) functionalized with a tetraethylene glycol oligomer and specificity of gene silencing using tumor cell lines that express reporter and injected them via various routes into rodents. Biodistribution and genes. We further studied scFv-mediated intracellular delivery of siRNAs excretion of APMS and gadolinium (Gd)-modified APMS were examined in that induce cell death. We performed cell viability studies in vitro for organs, peritoneal lavage fluids (PLF) and urine of normal mice and rats both epithelioid and sarcomatoid mesothelioma cell lines. Control non- over time. APMS was also functionalized with an antibody to mesothelin tumorigenic cells were included in the assay to assess off-target effects. (APMS-MB) or bovine serum albumin (BSA), a non-specific protein control, and in vivo tumor targeting was evaluated by inductively-coupled mass Results: Purified scFv-SRBM fusions retain tumor binding specificity of spectrometry and multifluorescence microscopy over a 6 day period. their parental scFvs. Like parental scFvs, these scFv-SRBMs are selectively Lastly, APMS-MB microparticles were loaded with doxorubicin (DOX), and internalized by both epithelial and sarcomatous mesothelioma cells but APMS-MB-DOX particles, DOX alone at the same concentrations, and not control non-tumorigenic cells. Using the reporter assay in vitro, we APMS-MB (no DOX) were injected intraperitoneally 3X weekly into SCID found that the scFv-SRBM-siRNA was able to significantly reduce reporter mice bearing human MMs. Tumor-bearing mice were also injected with protein expressions. The silencing effect is tumor-targeted as there is < 5% saline (controls). inhibition of reporter expression in the control non-tumorigenic cells. No reduction of reporter protein expression was observed when control siRNAs international mesothelioma interest group Results: APMS was primarily cleared via the urine over a 24 hr period, and were delivered. When cell death-inducing siRNAs were used in the viability small amounts were observed in liver, spleen and kidneys at 24 hr and 6 assay, we found that the scFv-SRBM-siRNA potently reduces viability of days. Neither inflammation nor necrosis was observed in major organs or both epithelial and sarcomatous mesothelioma cells in vitro. Furthermore, in PLF. Targeting with APMS-MB vs. APMS-BSA increased APMS uptake in the effect is specific to tumor but not control cells to which the scFv does mesenteric tumors at 6 days (p<0.05),Particles were observed in both MM not bind. No significant reduction in viability was observed for control cells and tumor-associated macrophages (TAM), and moved centrally into siRNAs or control non-binding scFvs. tumors over time. Approximately 10-12% of the initially injected amount was observed in both spheroid and mesenteric mesotheliomas at 6 days Conclusion: We have identified novel scFvs and SRBMs, and used them post-injection. Targeted therapy using APMS-MB-DOX was more effective to create multi-functional siRNA delivery vehicles that possess tumor than treatment with equivalent concentrations of DOX alone, resulting targeting, intracellular delivery, and siRNA binding functions. We have in the reduction of MM volume (p<0.05) primarily via inhibition of cell evaluated potency and specificity of our novel scFv-SRBM-siRNA in proliferation. Unlike mice injected with effective concentrations of DOX vitroon both epithelioid and sarcomatoid mesothelioma cell lines, and alone, mice receiving APMS-MB-DOX did not exhibit weight loss or organ demonstrated targeted gene silencing and selective tumor cell killing for toxicity after multiple injections. all subtypes of mesothelioma cells studied. Our approach is thus effective for tumor targeted intracellular siRNA delivery, a prerequisite for siRNA Conclusion: Our data suggest that localized delivery of APMS-MB into therapeutic development. We are performing in vivo systemic delivery the peritoneal or pleural cavity after encapsulation of drugs, plasmids studies to determine potency and toxicity of our novel internalizing or macromolecules is an effective strategy that should be tested in human antibody-based siRNA therapeutics for treating all subtypes of patients with MMs. Financial Support: This work was supported by the mesothelioma. Mesothelioma Applied Research Foundation (BTM); NCI STTR R41 CA126155 NIEHS T32 ES007122 (BTM for SLM, JMH, JLS); NCRR 1S10 RR08173-01A1 Disclosure: No significant relationships. and 1S10 RR01924B (DJT).

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 71 SESSION VIA NOVEL THERAPEUTICS: PRECLINICAL STUDIES Dox as a novel multimodality approach in the treatment of MM..Financial September 14, 2012 14:20-15:50 Support: This work was supported by the Mesothelioma Applied Research Foundation (BTM); NCI P01 CA11407 (BTM); NIEHS T32 ES07122 (BTM for VIA.5: INHIBITION OF MULTIPLE SIGNALING PATHWAYS BY SLM, JMH, JMM). We thank Dr. Harvey I. Pass (NYU) for the human MM cell VANDETANIB (ZD6474) INCREASES DOXORUBICIN-INDUCED lines. CELL TOXICITY IN HUMAN MESOTHELIOMA CELLS Disclosure: No significant relationships. Mutlay Sayan, Arti Shukla, Maximilian B. Macpherson, Sherrill L. Macura, Jedd M. Hillegass, Timothy N. Perkins, Joyce K. Thompson, Stacie L. Beuschel, Jill M. Miller, Brooke T. Mossman Pathology, University Of Vermont College Of Medicine, Burlington/VT/ SESSION VIA NOVEL THERAPEUTICS: PRECLINICAL STUDIES UNITED STATES OF AMERICA September 14, 2012 14:20-15:50

Background: Malignant mesothelioma (MM) is an aggressive malignancy VIA.6: A NOVEL THERAPY FOR MESOTHELIOMA USING HVJ-E with a poor prognosis. Therefore the need to develop novel and effective COMBINED WITH CHEMOTHERAPY. september 14, 2012 therapies is urgent. Vandetanib (Van) (ZD6474, ZACTIMA™) is a novel tyrosine kinase inhibitor (TKI) that originally was thought to inhibit Chunman Lee1, Nagako Sougawa1, Masao Sasai2, Yoshiki Sawa1, activation of the vascular endothelial growth factor receptor-2 (VEGFR-2), Yasufumi Kaneda3 as well as phosphorylation of the epidermal growth factor receptor (EGFR). 1Department Of Surgery, Osaka University, Suita/JAPAN, 2Med. Ctr. For WE have identified EGFR signaling as well as activation of extracellular Translational Research, Osaka University, Suita/JAPAN, 3Gene Therapy signal-related kinases (ERK1, ERK2, ERK5), Activator Protein-1 and cAMP Science, Osaka University, Suita/JAPAN response element binding protein (CREB) as critical signaling cascades and transcription factors that are activated by asbestos and elevated in Background: Immunotherapy for Malignant pleural mesothelioma (MPM) MMs (reviewed in Heintz NH, Janssen-Heininger, YMW, and Mossman, BT: is expected to be a breakthrough strategy, because it is suitable for the Asbestos, lung cancers, and mesotheliomas: from molecular approaches therapy of the disseminated lesions with minimal influence on normal to targeting tumor survival pathways. Am J Respir Cell Mol Biol 43:133-139, tissues. Various clinical studies of immunotherapy for MPM combined 2010). with chemotherapy have carried out, because these procedures lead to more effectiveness due to a large number of tumor antigens. We focused Methods: In human MM lines, HMESO and H2373, we evaluated whether the immunotherapy for MPM using the hemagglutinating virus of Japan Van had tumor cell killing efficacy both alone and in combination with envelope (HVJ-E). HVJ-E derived from inactivated replication-defective Doxorubicin (Dox) (Adriamycin) and the signaling cascades involved using Sendai virus enhances anti-tumor immunity through activation of effector cell toxicity assays and Western blot analysis. T cells and natural killer cells and inhibitory of regulatory T cells. The therapy using HVJ-E revealed highly effective eradication of murine colon Results: Van alone reduced total cell numbers in HMESO cells. Moreover, cancer, renal carcinoma, human prostate cancer, and human glioblastoma it synergistically increased the toxicity of Dox in vitro in both cell lines. in experimental medicine. We evaluated the therapeutic effectiveness of Van also caused the inhibition of Dox-induced phosphorylation of EGFR, HVJ-E combined with CDDP in human MPM bearing mice. ERKs, CREB, and protein kinase A (AKT) (see summary of results in Table 1 below). Table 1: A summary of the effects of Van and Dox alone and in Methods: Human MPM bearing mouse model: Two million cells of MSTO- combination on the phosphorylation of different cell signaling pathways as 211H were injected into the parietal pleura through the lower intercostal observed by Western blot analyses in MM cells space of 5 week-old female CB-17/SCID mice. Three microgram per weight (kg) of CDDP with 1,000 hemagglutination unit (HAU) of HVJ-E, or each material alone, was administrated into pleural cavity in the same maneuver on day8. One thousand HAU of HVJ-E was weekly administrated into

pERK1 pERK2 pERK5a pCREBa pAKT pEGFR international mesothelioma interest group subcutaneously space of mice belonging to HVJ-E combined with CDDP HMESO Van b group and HVJ-E group until mice death. We compared the survival time of 5µM control and test groups using the log-rank test. Dox       Results: Mean survival time of control group, HVJ-E (intrapleural injection 25µM at first time and weekly subcutaneous administration) group, CDDP (single Van       injection) group, and HVJ-E combined with CDDP group was 25.5, 38.8, 5uM 54.4, and 76.8 days, respectively. Significantly prolonged survival was + Dox seen for the HVJ-E combined with CDDP treatment group compared to the 25µMc other treatment groups (p < 0.01, log-rank test).

Conclusion: We demonstrated that a novel immunotherapy using H2373 Van HVJ-E combined with CDDP (single administration into pleural cavity 5µM and subsequent weekly subcutaneous administration of HVJ-E) showed Dox       enhanced antitumor effect against disseminated MPM and resulted in 100µM prolonged survival. These results suggest that HVJ-E combined with chemotherapy represents a potentially useful and convenient strategy for Van       MPM. 5µM + Dox Disclosure: No significant relationships. 100µM a pERK5 and pCREB present novel pathways first identified in this study b Arrows represent significant (p< 0.05) increases (up arrows) or decreases (down arrows) c All statistical comparisons with Van + Dox are with the Dox alone group. Comparisons involving individual agents are with the control group

Conclusion: Results suggest that a plethora of cell signaling pathways, apparently stimulated by Dox in a stress response are inhibited by Van in MM cells. Moreover, our results highlight the combined efficacy of Van and

iMig2012.org • Abstract Book 72 SESSION VIA NOVEL THERAPEUTICS: PRECLINICAL STUDIES September 14, 2012 14:20-15:50

VIA.7: INTRAVENOUS ADMINISTRATION OF FL118, A SURVIVIN/ MCL-1/XIAP/CIAP2 INHIBITOR, SHOWS HIGHLY EFFECTIVE INHIBITION OF MESOTHELIOMA CELL LINE-ESTABLISHED TUMOR GROWTH IN ANIMAL MODELS

Fengzhi Li, Shousong Cao, Xiang Ling Dept Of Pharmacology & Therapeutics, Roswell Park Cancer Institute, Buffalo/NY/UNITED STATES OF AMERICA

Background: We have identified and characterized a novel small chemical molecule (designated FL118). Our in vitro studies showed that FL118 selectively inhibits the expression of survivin, Mcl-1, XIAP and cIAP2, while september 14, 2012 showing no inhibitory effects on control genes. In an intraperitoneal (i.p.) route, the maximum tolerated dose (MTD) of FL118 was found to be about 1.5 mg/kg at a weekly x 4 schedule in a formulation of 0.05 mg/ml FL118, 5% DMSO, 20% Tween 80 and 75% saline, and at this in vivo experimental setting, FL118 showed superior antitumor activity in mouse models of human colon and head-&-neck tumor xenografts. However, the formulation used in the i.p. route is not intravenous (i.v.) compatible and also the i.p. route is not a common route in clinical practice with the exception of ovarian cancer.

Methods: Animal models and animal models of human mesothelioma cancer cell line-established tumor xenograft were used in these studies.

Results: Here, we report the use of a Tween 80-free formulation to test antitumor activity and toxicity (body weight loss) of FL118 via i.v. routes in animal models of human mesothelioma cancer cell line-established tumor xenograft. Important findings are summarized below. First, in contrast to that the MTD for FL118 in the i.p. formulation is 0.2 mg/kg in the daily x 5 schedule (5 does); 0.5 mg/kg in the every other day for three doses (q2 x 3, 3 doses); and 1.5 mg/kg in the weekly x 4 (4 doses) schedules, respectively, the MTD for FL118 in the Tween 80-free formulation increases 3-7 times without loss of FL118 antitumor activity via i.v. administration of the drug. Specifically, the MTD of FL118 in turn reached 1.5 mg/kg, 1.5-2 mg/kg and 5 mg/kg for daily x 5; q2 x 5 on day 0, 2, 4, 6, 8; and weekly x 4 schedules, respectively. Second, FL118, for the first time, showed highly effective to inhibit mesothelioma tumor growth and result in tumor regression in all three clinical compatible schedules, while the toxicity profile appears to be improved in comparison with the outcome from FL118 in the Tween 80-containing formulation. Specifically, our data international mesothelioma interest group showed that FL118 effectively inhibits MSTO-211H-derived tumor growth, induces tumor regression and even achieved a cure in a percentage of human mesothelioma xenograft, while the tumors in control mice without FL118 treatment reached the maximal size (~2000 mg/mm3) allowed by our protocol in less than five weeks. Additionally, FL118 also showed effective inhibition of NCI-H226-established tumor growth. These findings indicate that we developed a clinical suitable formula for FL118 i.v. administration. We also, for the first time, demonstrated that FL118 possesses superior antitumor activity in animal models of human mesothelioma cancer cell line-established tumor xenograft.

Conclusion: These studies would facilitate FL118 further development toward clinical trials and provide a hope to use FL118 for effective treatment of malignant human mesothelioma in clinical practice.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 73 Session VIB1 Local and Radiation Therapy September 14, 2012 14:20-15:00

SESSION VIB1 LOCAL AND RADIATION THERAPY SESSION VIB1 LOCAL AND RADIATION THERAPY September 14, 2012 14:20-15:00 September 14, 2012 14:20-15:04 september 14, 2012 VIB1.1: RADIATION PNEUMONECTOMY, A NOVEL OPTION THAT VIB1.2: FAILURE PATTERNS AFTER HEMITHORACIC INTENSITY- DELIVERS EFFECTIVE PALLIATION AND LOCAL CONTROL WITH MODULATED RADIATION THERAPY (IMRT) FOR MALIGNANT ACCEPTABLE TOXICITY IN PATIENTS WITH MALIGNANT PLEURAL PLEURAL MESOTHELIOMA (MPM) WITH TWO INTACT LUNGS MESOTHELIOMA Andreas Rimner1, Abraham J. Wu1, Ellen Yorke2, Amanda Mclane1, Malcolm Feigen1, Marita Lawlor1, Katheryn Churcher1, Kym Rykers1, Sze Marjorie G. Zauderer3, Kenneth E. Rosenzweig4, Valerie Rusch5, Lee M. Ting Lee2, Andrew M. Scott2, Chris Hamilton1 Krug6 1Radiation Oncology Center, Austin Health, Heidelberg West/VIC/ 1Department Of Radiation Oncology, Memorial Sloan-Kettering Cancer AUSTRALIA, 2Centre For Pet, Ludwig Institute For Cancer Research, Austin Center, New York/UNITED STATES OF AMERICA, 2Department Of Medical Health, VIC/AUSTRALIA Physics, Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF AMERICA, 3Department Of Medicine, Memorial Sloan-Kettering Background: Malignant pleural mesothelioma is an incurable disease Cancer Center, New York/NY/UNITED STATES OF AMERICA, 4Department that commonly relapses after chemotherapy and conventional Of Radiation Oncology, Mount Sinai Medical Center, New York/NY/ radiotherapy. Surgeons are increasingly reluctant to perform extrapleural UNITED STATES OF AMERICA, 5Department Of Thoracic Surgery, Memorial pneumonectomies (EPP) as there is high postoperative morbidity and Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF survival rates are low. We developed a program utilizing new technologies AMERICA, 6Department Of Medicine, Thoracic Oncology Service, Memorial to optimize palliation with high-dose radiotherapy for patients with locally Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF AMERICA advanced disease who had not had an EPP. Background: We recently reported our initial experience with definitive Methods: In 2003 Stevens et al demonstrated that hemithoracic intensity- or adjuvant hemithoracic pleural IMRT for MPM patients with two intact modulated radiotherapy (IMRT) provided effective adjuvant therapy to lungs. Given the steep dose gradient with IMRT, accurate target delineation minimize locoregional relapses after EPP. We modified their technique is critical. Here, we present a detailed failure pattern analysis in relation to for patients with unresected mesothelioma localized to the ipsilateral target delineation and radiation fields. hemithorax, using 18F-FDG PET scans to outline tumor masses. Previous surgical procedures and chemotherapy were documented. Acute and late Methods: We retrospectively reviewed the RT plans and failure patterns toxicities were graded using CTCAE version 4. of 64 patients with MPM treated with definitive or adjuvant pleural IMRT between 04/2005 and 05/2012. All patients had a PET scan available for Results: Over 9 years, 51 patients received RT to 45-60 Gy, in 16 using target delineation. The median dose was 4680cGy in 26 fractions (range

3D-conformal radiotherapy and 35 with IMRT. Patients were aged 45-76 3960cGy – 5040cGy). Three patients received an integrated boost to gross international mesothelioma interest group years, mainly males (86%) with right-sided (65%) mesotheliomas. 16% disease to a maximum dose of 5994cGy. Treatment plans were optimized had non-epithelioid histologies and 82% had clinical stage III/IV disease. to keep the mean lung dose <21 Gy. Failures were categorized as in-field 24 had a prior pleurectomy/decortication, 21 a pleurodesis and 6 biopsy local failures (LF) (within the 90% isodose line (IDL)), marginal failures only. Five had planned trimodality therapy and 24 had progressed post- (<90% and ≥50% IDL) and out-of-field failures (outside the 50% IDL). The chemotherapy. At median followup of 9.5 months (range 4-90 months), median age at diagnosis was 66 years (range 42 to 82 years). Forty-nine there were no deaths from radiation toxicity. 82 followup PET/CT scans patients had epithelioid histology, and 15 patients had sarcomatoid or were reviewed in addition to routine CT scans. 8 patients recurred biphasic MPM. Thirty-four patients had right-sided and 30 patients left- within the planning target volumes (PTV), all with concurrent metastases sided MPM. Fifty percent presented with early-stage (clinical stage I/II) and documented in unirradiated sites. 31 recurred only outside the PTV. 16 50% with advanced-stage MPM (clinical stage III/IV). Thirty-eight patients remain alive, 10 disease-free. There were no significant acute radiation underwent a partial or complete pleurectomy/decortication (P/D), while toxicities and no radiation pneumonitis > grade 3 in the absence of disease 26 patients were technically or medically unresectable. Fifty-eight patients in the contralateral lung. (91%) received neoadjuvant or adjuvant platinum/pemetrexed-based chemotherapy. Conclusion: High-dose hemithoracic RT is recommended to improve locoregional control post-EPP, and should now be considered for selected Results: With a median followup of 16 months from diagnosis and mesothelioma patients whose lungs remain intact. Advanced radiotherapy 7.4 months from the end of last treatment, the median in-field local techniques will minimize normal tissue toxicities and provide long-term failure-free survival was 8.9 months. Thirty-nine in-field LFs (61%) palliation, particularly in those patients whose mesothelioma does not were found, with 29 failures occurring in sites of previous gross disease spread beyond the hemithorax. and 10 in a previously grossly uninvolved site. LF was the first site of failure in 19 patients (30%). The only factors associated with higher Disclosure: No significant relationships. local failure-free survival were left-sided MPM (p=0.03) and a trend for patients who underwent surgical resection (p=0.07). Response to chemotherapy or RT dose was not significantly associated with local failures. There were 13 marginal failures (20%). Increasing experience over time was correlated with decreasing marginal failures (p=0.03). On review of the imaging studies at time of target delineation, in 6 cases we identified an abnormality that had not been included in the treatment volume. Four cases represented supradiaphragmatic lymph nodes in the costomediastinal recess, and two cases were located along the crus in the costodiaphragmatic recess. Two of these patients experienced an isolated

iMig2012.org • Abstract Book 74 local failure. Seven patients (11%) failed in the fissures, but only one patient had an isolated failure in the fissure without pleural failures elsewhere. Twenty-one patients (33%) had out-of-field failures in mediastinal lymph nodes, but none were isolated elective nodal failures.

Conclusion: After pleural IMRT most LFs occur in sites of previous gross disease. Thus, surgical resection of sites of gross disease remains important. Increasing experience and improvements in target delineation may decrease the incidence of marginal failures.

Disclosure: No significant relationships. september 14, 2012 international mesothelioma interest group

iMig2012.org • Abstract Book 75 Session VIB2 Asbestos Epidemiology September 14, 2012 15:00-15:50

SESSION VIB2 ASBESTOS EPIDEMIOLOGY which further their own purposes. Such sponsorship should no longer be September 14, 2012 15:00-15:50 accepted by scientific assemblies. september 14, 2012 VIB2.1: MISREPRESENTATION OF HISTORICAL FIBER RESEARCH: Disclosure: No significant relationships. AN INTEVIEW-BASED STUDY FOR RETROSPECTIVE KNOWLEDGE TRANSLATION OF EPIDEMIOLOGICAL INQUIRY.

Bruce W. Case Pathology Epidemiology And Occupational Health, McGill University, Montreal/CANADA

Background: Epidemiological research linking disease outcomes and exposures to fibers dates from the late 1950’s. There is a poor knowledge and comprehension of this research and its vital role among current public health professionals and stakeholders.

Methods: The most prominent researchers from the beginnings of this research were interviewed in person on three continents in 2004 and 2005. Where the researchers had died, information was directly obtained from similar interviews of relatives and co-workers. Interviews were free-form and unstructured. In the period following interviews additional information was obtained from publicly available documents, documents provided by interviewees, and other sources. Court documents and such sources as newspaper or magazine articles were not considered due to the possibility of bias. Information was obtained from Richard Doll, Julian Peto, Dr. Margaret Wagner, Dr. Corbett McDonald, Dr. Arthur Langer, Dr. G. Berry, Dr. G. Hillerdal, Dr. P. Sebastien, and many others involved in the original works.

Results: Information obtained conflicted in many ways from “conventional wisdom”. Original researchers conveyed many regrets about the ways in

which past research has been received and acted (or not acted) upon. international mesothelioma interest group Researchers were particularly concerned with misrepresentations of their lives and work, and with a general lack of knowledge of what was known in the past in relation to what is believed now. There was an evident lack of knowledge translation to the population in general and to stakeholder groups in particular. Misrepresentation and character assassination were frequent complaints by researchers on all “sides”. This presentation will offer examples of the ways in which the pioneer researchers and seminal pieces of research regarding exposure and disease, particularly in relation to mesothelioma, have been misunderstood, and in some instances, used by for-profit agents or tainted by ideological bias.

Conclusion: 1. There is poor knowledge among current health practitioners and researchers about historical research development for the epidemiology of asbestos-related diseases. 2. This is partly generational, and can be addressed through “retrospective knowledge translation”, conveyed to health professionals and through them to stakeholders and the broader public. 3. Lack of knowledge has led to mistrust of epidemiological research for fiber-related research, due in part to high numbers of publications with varying methodology and motivation over more than a half century. 4. There are three principal reasons for the above problems: (a) Ideological bias and a system of “fixed beliefs”. These can be alleviated through a decrease in mutually exclusive activities of stakeholders and increased cooperation. (b) Lack of knowledge of the facts and results of past research. This is the most difficult problem to address. “Retrospective knowledge translation” activities such as this study are needed. (c) Financial bias, particularly in American asbestos litigation, a for-profit activity not constrained by normal academic requirements and related in part to the contingency-fee basis of such activity. This extends to funding of scientific assemblies, including those of iMig, by law firms with financial interests in the promotion of ideas

iMig2012.org • Abstract Book 76 Session VIC1 Gene Expression Profiling September 14, 2012 14:20-15:00

SESSION VIC1 GENE EXPRESSION PROFILING SESSION VIC1 GENE EXPRESSION PROFILING September 14, 2012 14:20-15:00 September 14, 2012 14:20-15:00 september 14, 2012 VIC1.2: COMPARATIVE GENE EXPRESSION ANALYSIS OF EX- VIC1.3: BIOINFORMATICS ANALYSIS OF THORACIC VIVO FINE NEEDLE ASPIRATION BIOPSIES AND MATCHED SOLID MALIGNANCIES MICROARRAYS IDENTIFIES NOVEL GENE TISSUES IN MALIGNANT PLEURAL MESOTHELIOMA FUNCTIONS IN MALIGNANT PLEURAL MESOTHELIOMA

Assunta De Rienzo1, Yaoyu E. Wang2, William G. Richards1, David J. Assunta De Rienzo1, Yaoyu E. Wang2, Beow Y. Yeap3, David J. Sugarbaker1, Raphael Bueno1 Sugarbaker1, Raphael Bueno1 1Division Of Thoracic Surgery, Brigham And Women’s Hospital And Harvard 1Division Of Thoracic Surgery, Brigham And Women’s Hospital, Boston/ Medical School, Boston/MA/UNITED STATES OF AMERICA, 2Center For MA/UNITED STATES OF AMERICA, 2Center For Cancer Computational Cancer Computational Biology, Dana-Farber Cancer Institute, Boston/MA/ Biology, Dana-Farber Cancer Institute, Boston/MA/UNITED STATES OF UNITED STATES OF AMERICA AMERICA, 3Department Of Medicine, Massachusetts General Hospital And Harvard Medical School, Boston/MA/UNITED STATES OF AMERICA Background: FNA biopsy, whether image-guided or by direct palpation, is a useful diagnostic minimally invasive method for patients with Background: Malignant pleural mesothelioma (MPM) is an aggressive presumed neoplasms. However, the accuracy, specificity and sensitivity malignancy arising from the mesothelial cells of the pleura. The expected of the cytological interpretation of FNA may be limited in a variety of median survival of patients diagnosed with MPM is between 4 and 12 circumstances such as target size, tumor type, accessibility etc. One major months. Consequently, there is an urgent need for a better understanding limitation is the need for intact recognizable tumor cells and architecture of the major molecular pathways of MPM to establish diagnostic, for definitive histological typing and diagnosis. Previously, we have shown therapeutic and prevention methods. To search for insights via differential that FNA biopsies are a feasible tool to perform molecular tests in MPM and gene expression into pathways uniquely relevant in MPM in comparison to lung cancer, however, a systematic comparative analysis of expression data other related cancers, we performed bioinformatics analysis of large data between FNAs and matched solid tumors in thoracic malignancies has not set of expression array data. been conducted so far. Methods: Expression array analysis was carried out on 112 specimens Methods: We performed whole genome expression of 192 samples including 39 MPMs, and a spectrum of all the other malignancies and including 164 Mesothelioma (MPM) solid tumors, 8 matched epitheliod benign tissues in the differential diagnosis of MPM using Illumina whole- MPM ex-vivo FNAs, 8 adenocarcinoma ex-vivo FNAs, 8 squamous cell genome microarrays. For clustering and functional enrichment analysis, carcinoma ex-vivo FNAs, and 4 controls using the Affymetrix Human the arrays were normalized by quantile normalization using Bioconductor, Gene 1.1 ST Array. The samples that passed quality control were quantile and differentially expressed probes were identified by linear model using

normalized to reduce array specific biases. To determine whether different the LIMMA package. Probes with q-value of <0.01 and log fold change international mesothelioma interest group sample sets were grouped together or separated in different branches, >1.5 obtained using multiple comparison were included in the analysis. significant differentially expressed genes between epithelioid and Hierarchical clustering was performed with Euclidean distance in R. The sarcomatoid MPMs were identified using linear model (q<0.01 and fold 167 probes identified in this analysis were further analyzed in 36 MPM change>2), and their gene expression profiles were employed for clustering samples to detect probes differentially expressed between epitheliod and analysis. Finally, to determine whether the MPM FNA gene expression sarcomatoid MPMs using the same methods. The overall survival of 6 genes was comparable to their matched solid tissues, we performed principle up-regulated in the sarcomatoid group was estimated by the Kaplan–Meier component analysis and compared Euclidean distances between each MPM method. FNA and its matching tumor tissue with randomly chosen samples. Results: One hundred and sixty-seven probes, corresponding to 156 Results: Whole array analysis was performed and 4 outliers in the MPM unique genes, were identified as differentially expressed (pvalue < 0.01) solid tumors group were excluded for further analysis. Forty-two probes between MPM and other tumor types. These probes were employed to were found differentially expressed between epithelioid and sarcomatoid perform hierarchical clustering analysis and a cluster dendrogram showing MPM samples. Hierarchical clustering using these probes resulted in a tree two major branches was obtained. Thirty-six MPM samples clustered in a structure where all the MPM FNAs were grouped with the epithelioid MPM single branch. The epithelioid and the sarcomatoid MPMs were in distinct solid samples showing that the matched samples grouped in the same groups indicating that that the selected gene expression profile can branch. In addition, the principle components analysis showed that all segregate the two groups. Gene set enrichment analysis was performed on MPM FNAs clustered with the epithelioid MPM, whereas the sarcomatoid the selected genes and 45 pathways were significantly enriched (FDR < 20) MPM and lung cancer FNAs clustered into two different groups. The in the MPM group, and were classified into four main groups: extracellular Euclidean distance between each FNA biopsy and its matched tumor tissue organization, development, response to endogenous, mechanical, or was significantly smaller compare to the distance between randomly hormonal stimuli, and immune response. Focused analysis within the MPM selected tumor tissues suggesting that the matching samples exhibit group identified 51 probes differentially expressed between the epitheliod similar gene expression profile. Finally, only 11 probes corresponding to and sarcomatoid groups with 45 up-regulated in epithelioid MPM, and 9 unique genes were found differentially expressed between MPM FNAs 6 probes up-regulated in the sarcomatoid subtype. Interestingly, all the and matched normal tissues indicating that the expression profile of the proteins corresponding to the probes up-regulated in the sarcomatoid matched pair is comparable. MPM are localized in the extracellular space or on the plasma membrane and are related to the epithelial-to-mesenchymal transition process. In Conclusion: Our study shows that the ex-vivo FNA biopsies have gene an independent set of 129 epitheliod MPMs, 4 of the 6 genes showed expression comparable to the corresponding solid tissues, suggesting that expression associated with survival (p < 0.05). FNA biopsies from thoracic malignancies can be used for molecular studies. Conclusion: Our work provides molecular evidences of genes and Disclosure: No significant relationships.

iMig2012.org • Abstract Book 77 pathways specifically activated in MPM. Insights into the molecular basis of MPM may facilitate a personalized treatment approach involving early identification of poor prognostic indicators that may reduce the heterogeneity of the clinical response and lead to more focus treatments.

Disclosure: No significant relationships. september 14, 2012 international mesothelioma interest group

iMig2012.org • Abstract Book 78 Session VIC2 Novel Therapeutics: Preclinical Studies September 14, 2012 15:00-15:50

SESSION VIC2 NOVEL THERAPEUTICS: PRECLINICAL STUDIES September 14, 2012 15:00-15:50 september 14, 2012 VIC2.1: MITHRAMYCIN IS A NOVEL THERAPEUTIC AGENT FOR MALIGNANT PLEURAL MESOTHELIOMA

Scott M. Atay1, Mahadev Rao1, Xinmin Li2, Trevor Upham3, Patricia Fetsch4, Markku Miettinen4, Suzanne Inchauste3, Julie Hong3, Mary Zhang3, Sichuan Xi3, Assunta De Rienzo5, Raphael Bueno6, David S. Schrump3 1Thoracic Oncology Section, Surgery Branch, National Cancer Institute, Nih, Bethesda/UNITED STATES OF AMERICA, 2Pathology And Laboratory Medicine, Clinical Microarray Core, David Geffen School Of Medicine At The University Of California, Los Angeles, Los Angeles/UNITED STATES OF AMERICA, 3Thoracic Oncology Section, Surgery Branch, Center For Cancer Research, National Cancer Institute, Nih, Bethesda/UNITED STATES OF AMERICA, 4Laboratory Of Pathology, Center For Cancer Research, National Cancer Institute, Nih, Bethesda/UNITED STATES OF AMERICA, 5Division Of Thoracic Surgery, Brigham And Women’s Hospital, Harvard Medical School, Boston/UNITED STATES OF AMERICA, 6Division Of Thoracic Surgery, Brigham And Women’s Hospital And Harvard Medical School, Boston/MA/ UNITED STATES OF AMERICA Figure 1. Mithramycin inhibits tumor growth and mediates tumor Results: Pleural mesothelioma lines exhibited significantly higher Sp1 regression in established subcutaneous xenografts. Size of tumors in mice expression levels relative to cultured normal mesothelial cells. IHC analysis treated with MM were significantly smaller than controls (saline) in 2 MPM demonstrated over-expression of Sp1 in 73% (14/19) of MPM specimens cell lines ( * P<0.02 control vs. 1mg/kg and 2mg/kg). relative to normal pleura controls (n=3). Knock-down of Sp1 as well as MM treatment significantly inhibited proliferation and clonogenicity of MPM Disclosure: No significant relationships. cells. Intraperitoneal MM administered at either 1 mg/kg or 2 mg/kg every other day mediated dose dependent growth inhibition and regression of established subcutaneous MPM xenografts (p <0.02 relative to saline controls; figure 1). These effects coincided with dramatic alterations in SESSION VIC2 NOVEL THERAPEUTICS: PRECLINICAL STUDIES international mesothelioma interest group global gene expression profiles; a gene expression signature corresponding September 14, 2012 15:00-15:50 with response to MM was identified. VIC2.2: DRUG SENSITIVITY SCREENING OF SHORT-TERM PRIMARY TUMOR CULTURES OF MALIGNANT PLEURAL MESOTHELIOMA

Josine Quispel-Janssen1, Laurel Schunselaar2, Jacques Neefjes2, Paul Baas1 1Thoracic Oncology, Nki-Avl, Amsterdam/NETHERLANDS, 2Cell Biology, Nki-Avl, Amsterdam/NETHERLANDS

Background: Malignant pleural mesothelioma (MPM) is a cancer with a poor prognosis. Only 40% of patients respond to combination chemotherapy (cisplatin and pemetrexed) in first line setting and survival benefit is limited. No predictive markers are available yet to identify patients that may benefit from chemotherapy. Assessing in vitro chemotherapeutic drug resistance has been demonstrated to be feasible using MPM resection specimens in a commercial assay. This assay included only 3 different drugs, excluding pemetrexed and combinations of drugs. However, obtaining resection specimens is not feasible for all patients. Furthermore, a chemosensitivity and resistance assay should at least include the standard chemotherapy regimen and be able to evaluate combinations of drugs. Our goal was to develop robust protocols to use tumor cells from pleural fluid of MPM patients for chemosensitivity and resistance testing.

Methods: Cells are isolated from pleural fluid, drawn from patients with MPM for symptom relief. The cells are cultured under low oxygen conditions for a period of 4 weeks and grow in adherent monolayers. Cultures contain

iMig2012.org • Abstract Book 79 both tumor cells and stroma cells. Cell morphology, viability and tumor this cancer. percentage are assessed by cytopathological staining using Giemsa, anti-pankeratine to check for epithelial phenotype and Ki-67 to assess Methods: We have extended these studies to test whether vitamin D, non- proliferation rate at each passage. During week two and three of culture, steroidal anti-inflammatories, statins and some other candidate diets could drug sensitivity was measured. Cells are plated and incubated with an 8 alter the pattern of disease in the MexTAg model. Supplemented diets were point concentration range of 5 single drugs and 2 two-drug combinations provided at levels based on published data and began 2 weeks prior to for 48 hours. Cell viability is determined by the Cell Titer Blue assay. Each asbestos exposure in order to maximize our chance of detecting a benefit. concentration point is measured in triplo and a biological duplo experiment is performed to check reproducibility. Results: We found Vitamin D, apigenin, lupin, linseed and diallyl trisulphide (garlic derivative) did not alter the profile of disease. Aspirin was tested Results: Fourteen out of 20 isolations (70%) resulted in successful cultures. at low and high levels (6 or 25 mg/kg/day). Again, this did not affect the Ninety percent of patients had epitheloid subtype and 10% mixed subtype. rate of disease development or progression. Similarly, three different Tumor percentage varied between 40-70% at isolation and increased up concentrations of statins in diets (10, 20 and 40 mg/kg/day) had no effect to 80-90% after the first passage. Percentage Ki-67 positive cells varied on the development of disease. between 15% en 60% between individual cultures. Anti-pankeratine september 14, 2012 staining was positive at all passages indicating that tumor cells maintain Conclusion: In conclusion, we think it is unlikely that antioxidants, anti- their epitheloid phenotype and no epithelial-mesenchymal transition inflammatories or other nutrient-specific diets will moderate the rate of occurs during culture. Drugs tested for were cisplatin or carboplatin, mesothelioma in asbestos exposed populations. pemetrexed, gemcitabine, vinorelbine, oxaliplatin and a combination of cisplatin and pemetrexed and of oxaliplatin and gemcitabine. Drug Disclosure: No significant relationships. screening was performed in seven different primary tumor cultures and demonstrated to be reproducible. Dose-response curves showed different sensitivity to the various drugs for the different primary tumor cultures. One patient had drainage of pleural fluid at two different time points and SESSION VIC2 NOVEL THERAPEUTICS: PRECLINICAL STUDIES tumor cells were cultured twice. Results of both drug screens were similar. September 14, 2012 15:00-15:50 Two patients that demonstrated resistance to the combination of cisplatin and pemetrexed in vitro, had progressive disease after three courses of this VIC2.4: INTRATUMORAL DRUG DELIVERY VIA EXPANSILE chemotherapy regimen in first line treatment. NANOPARTICLES FOR HUMAN MALIGNANT PLEURAL MESOTHELIOMA IN VITRO AND IN IN VIVO MODELS Conclusion: Short-term primary tumor cultures from pleural fluid of mesothelioma patients can be generated with high succes rate. A Rong Liu1, Denis M. Gilmore1, Barry C. Gibney1, Aaron Colby2, Morgan D. chemosensitivity and resistance assay with these short-term primary tumor Schulz1, Robert F. Padera3, Mark W. Grinstaff2, Yolonda L. Colson1 cultures is feasible. These cultures may be suitable for pharmacogenomic 1Surgery, Brigham And Women’s Hospital, Boston/MA/UNITED STATES profiling using high-troughput drug screens. OF AMERICA, 2Chemistry And Biomedical Engineering, Boston University, Boston/MA/UNITED STATES OF AMERICA, 3Pathology, Brigham And Disclosure: No significant relationships. Women’s Hospital, Boston/MA/UNITED STATES OF AMERICA

Background: Despite aggressive multimodality therapy, including pleurectomy or extrapleural pneumonectomy, overall survival for SESSION VIC2 NOVEL THERAPEUTICS: PRECLINICAL STUDIES patients with malignant pleural mesothelioma (MPM) is severely limited September 14, 2012 15:00-15:50 by a high incidence of locally recurrent disease in the chest and/or abdomen. Expansile nanoparticles loaded with paclitaxel (Pax-eNP) VIC2.3: STRATEGIES FOR THE PREVENTION OF MESOTHELIOMA have demonstrated superior efficacy in the prevention of intraperitoneal IN MEXTAG MICE (IP) growth of a human biphasic mesothelioma cell line (MSTO-211H) international mesothelioma interest group when injected on the same day in a xenogeneic nude mouse model. We Cleo Robinson, Samantha Woo, Kelly Martinovich, Amy Walsh, Anna K. hypothesized that increased efficacy was due to increased contact of Nowak, Richard Lake drug and tumor cells. To investigate this hypothesis, we assessed both in School Of Medicine And Pharmacology, National Centre For Asbestos vitro cellular uptake and tumor cytotoxicity of eNP against MSTO-211H Related Diseases, University Of Western Australia, Perth/WA/AUSTRALIA and human mesothelioma cells isolated from a malignant pleural effusion and in vivoeNP localization within a xenogeneic mouse model of established Background: Current treatments for mesothelioma typically increase pleural mesothelioma. median survival by a matter of months. Progress in treatment has been hampered by lack of a suitable small animal model, which could Methods: Intracellular uptake of eNP was demonstrated using confocal guide clinical advances, given limited numbers of patients eligible for microscopy and quantitated with flowcytometric analysis. Pax-eNP clinical trials. To this end we recently developed a transgenic mouse cytotoxicity of tumor cells was calculated via the standard MTT cytotoxicity model of mesothelioma in which the viral oncogene, SV40TAg (TAg) is assay. In vivo uptake was assessed in athymic mice, with the intrapleural directed to mesothelial cells by use of the cell type specific mesothelin injection of rhodamine-labeled eNP (rho-eNP) in the left chest, two weeks promoter. MexTAg mice develop mesothelioma rapidly and uniformly, after establishment of luciferase-transfected human mesothelioma tumor but onlyfollowing exposure to the natural carcinogen, asbestos. The (MSTO-211-luc, 1x106). Intratumoral localization was assessed four days model closely mimics the human disease and is thus ideal for both rapid later using UV imaging and confocal imaging. analysis of novel therapeutic studies and for investigating factors that might act synergistically with asbestos to cause disease. Since all MexTAg Results: Nanoparticle-mediated intracellular drug delivery was mice develop mesothelioma following asbestos exposure the model is confirmed in vitro with colocalization of Rho-eNP and Oregon-green Pax highly suitable for early intervention and cancer prevention studies. An within tumor cells by 4 hours. Dose-dependent cytotoxicity of pax-eNP, effective cancer prevention strategy for the millions of people who have but not unloaded eNP, was presented in vitro against human mesothelioma been exposed to asbestos could have enormous benefit worldwide. cells derived from a patient effusion or against the MSTO cell line. UV and Epidemiological evidence indicates that supplementation with some confocal imaging of intrathoracic MPM in vivo demonstrated that Rho-eNP dietary factors or use of common drugs such as statins and non-steroidal localize with high affinity to intrapleural tumor nodules in situ (Figure 1), anti-inflammatory drugs is associated with a lower incidence of cancer. and deliver paclitaxel directly into tumor in situ. We previously reported that dietary supplementation with a number of antioxidants did not alter the time to develop disease nor overall survival, Conclusion: Pax-eNP exhibit time dependent uptake and dose dependent despite the widely accepted hypothesis that asbestos catalyzed production cytotoxicity against human mesothelioma cells, including cells within a of reactive oxygen and nitrogen species contribute to the development of malignant effusion. Similarly, drug-loaded eNP demonstrate higher affinity

iMig2012.org • Abstract Book 80 to tumor tissues in vivo than to normal tissues, resulting in increased contact time between drug-eluting eNP and sites of tumor in vivo. Future studies will assess the efficacy of intrapleural delivery of pax-eNP for the control of local recurrence following pneumectomy for the treatment of MPM.

Disclosure: No significant relationships. september 14, 2012 international mesothelioma interest group

iMig2012.org • Abstract Book 81 Poster Session 1 Surgery September 12, 2012 11:30-12:30

POSTER SESSION 1 September 12, 2012 11:30-12:30 Background: Main principle of surgery in malignant pleural mesothelioma (MPM) is macroscopic complete resection. Adjuvant treatments are P1.01: IS THERE LIFE AFTER MARS? DOES EXTRAPLEURAL utilized to control microscopic invasions or micrometastasis. The impact PNEUMONECTOMY STILL HAVE A ROLE IN THE MANAGEMENT of microscopic margin positivity on survival and recurrence is not known OF MALIGNANT PLEURAL MESOTHELIOMA - A 13 YEARS SINGLE and thus we analyzed this issue in patients who underwent extrapleural

CENTRE EXPERIENCE. pneumonectomy (EPP) for epithelial MPM. poster sessions | september 12

Alex Cale1, Suhail Qadri2, Michael Cowen3, Andrezej Wieczorek2, Methods: The patients with epithelial MPM who underwent EPP over a Michael J. Lind4 period of 7 years were included in the study. Patients with non-epithelilal 1Cardiothoracic Surgery, Castle Hill Hospital, 5jq/UNITED histology (n=5), postoperative mortalities (n=5), patients who died of other KINGDOM, 2Castle Hill Hosp/UNITED KINGDOM, 3Castle Hill Hospital/ causes (n=5) were excluded from the study. Post surgical EPP specimens UNITED KINGDOM, 4Postgraduate Medical Institute, University Of Hull, were sampled from >20 sites at mediastinal, apical, lateral costal and Hull/UNITED KINGDOM costodiaphragmatic areas. Pericardial and transdiaphragmatic invasion were evaluated as well. If no microscopic invasion was noted in the areas Background: The MARS trial has shown no survival benefit of extrapleural above, the resection was accepted as a microscopic complete resection. pneumonectomy for the treament of malignant mesothelioma. We aim to Microscopic margin positivity in more than one area was evaluated as a present our results which are in contrast to the MARS trial results. prognostic factor. Recurrence rate and sites of recurrences were recorded. Data was evaluated using uni- and multivariate and Kaplan Meier Survival Methods: Patients who underwent extrapleural pneumonectomy for analysis. malignant mesothelioma between march 1999 and April 2011 were analysed retrospectively and their survival was observed until May 2012. Results: A total of 25 patients (Average age 52 [34-70], 11 females) were Risk was calculated by using thoracoscore. evaluable. Macroscopic complete resection was achieved in all patients. 23 patients completed adjuvant hemithoracic irradiation and 3 cycles of platin Results: Thirty patients underwent extrapleural pneumonectomy during based chemotherapy. Median survival in the whole group was 25,7 months this period. Median age was 59±8 years, with 29 male and 1 female. The (15% at 5-year). 12 patients had extrapleural lymph node metastasis. mean thoracoscore was 7.9±2.5. There was no in-hospital or 30 day Microscopic complete resection was achieved in 5 patients. Microscopic mortality. Overall mean survival was 25 months; increasing to 38 months mediastinal, apical, lateral costal, costodiaphragmatic, pericardial and for those that completed tri-modality treatment. Eight patients survived transdiaphragmatic invasion was present in 9, 9, 14, 3, 3 and 2 patients over 4 years and two over 5 years. Survival was significantly higher in respectively. Extrapleural lymph node metastasis (p=0,05) and microscopic epitheliod versus biphasic mesothelioma, right versus left EPP, age below margin positivity in more than one area (p=0,04) were significant factors 65 years, and with no N2 disease. 6, 12 and 18 month survival was 79%, in univariate analysis. In multivariate analysis, only microscopic margin

62.5% and 54% in comparison to 65%, 52%, and 34% in the MARS trial. positivity in more than one area (p=0,046) was the significant factor. international mesothelioma interest group Peritoneal or abdominal wall recurrence (8 patients) was common in Conclusion: Epitheliod malignant mesothelioma, right EPP, negative extral patients with microscopic complete resection or positivity only in one area pleural lymph nodes and age under 65 years are associated with prologed (n=12). survival. The UK National Thoracic Surgery Database report states that 12 EPPs were carried out in the UK over the last 3 years with no operative Conclusion: Macroscopic complete resection may be the goal of MPM mortality. Extrapleural pneumonectomy still has a role in the management surgery, but microscopic complete resection appears to influence survival of malignant mesothelioma in selected patients by experienced surgeons, significantly. Thus every effort should be made to achieve complete and in the context of a tri-modality therapy programme. resection margins in patients with epithelial MPM.

Disclosure: No significant relationships. Disclosure: No significant relationships.

POSTER SESSION 1 September 12, 2012 11:30-12:30 POSTER SESSION 1 September 12, 2012 11:30-12:30

P1.03: MICROSCOPIC MARGIN POSITIVITY IN MORE THAN ONE P1.05: OUTCOME OF CONSECUTIVE 61 CASES OF INTENT- AREA IN THE THORACIC CAVITY IS A POOR PROGNOSTIC FACTOR TO-TREAT WITH EXTRAPLEURAL PNEUMONECTOMY FOR FOLLOWING EXTRAPLEURAL PNEUMONECTOMY IN PATIENTS RESECTABLE MALIGNANT PLEURAL MESOTHELIOMA WITH EPITHELIAL MALIGNANT PLEURAL MESOTHELIOMA Seiki Hasegawa1, Fumihiro Tanaka2, Nobuyuki Kondo1, Yoshitomo 3 1 1 1 Onur Ermerak1, Fulden Yumuk2, Hale Basak Ozkok3, Rengin Ahiskali4, Okumura , Seiji Matsumoto , Teruhisa Takuwa , Masaki Hashimoto , 1 1 3 1 Muzaffer Metintas5, Hasan F. Batirel1 Hayato Orui , Ayumi Kuroda , Shunichi Fukuda , Kazue Yoneda , Noriaki 4 5 6 7 1Thoracic Surgery, Marmara University Hospital, Istanbul/TURKEY, 2Division Tsubota , Norihiko Kamikonya , Kazuya Fukuoka , Ikuko Torii , Tohru 7 6 Of Medical Oncology, Department Of Internal Medicine, Marmara Tsujimura , Takashi Nakano 1 2 University Hospital, Istanbul/TURKEY, 3Department Of Radiation Oncology, Thoracic Surgery, Hyogo College Of Medicine, Nishinomiya/JAPAN, Second Acibadem University, Istanbul/TURKEY, 4Pathology, Marmara University Department Of Surgery, University Of Occupational And Environmental 3 Hospital/TURKEY, 5Department Of Chest Diseases, Eskisehir Osmangazi Health/JAPAN, Department Of Thoracic Surgery, Itami City Hospital/ 4 5 University Medical Faculty/TURKEY JAPAN, Thoracic Oncology, Hyogo College Of Medicine/JAPAN, Radiation Oncology, Hyogo College Of Medicine/JAPAN, 6Respiratory Medicine, Hyogo College Of Medicine/JAPAN, 7Pathology, Hyogo College Of Medicine/JAPAN

iMig2012.org • Abstract Book 82 Background: The role of surgery in patients with resectable malignant POSTER SESSION 1 September 12, 2012 11:30-12:30 pleural mesothelioma (MPM) remains to be a matter to debate. Only a prospective randomized study comparing surgery versus no-surgery can P1.06: CYTOREDUCTIVE SURGERY AND THE IMMUNE SYSTEM— provide an answer but is almost unrealistic. The second best way is an A MECHANISM TO MAXIMIZE ADJUVANT IMMUNOTHERAPY intent-to-treat based analysis on prospectively registered patients. APPROACHES

Methods: We reviewed all patients in a prospective database of Jarrod D. Predina1, Brendan Judy1, Veena Kapoor1, Zvi G. Fridlender2, extrapleural pneumonectomy (EPP) program of Hyogo College of Medicine Steven M. Albelda2, Sunil Singhal1 between July 2004 and March 2012. Patients were eligibile if they had 1Division Of Thoracic Surgery, University Of Pennsylvania School Of a histologically confirmed MPM of any type, clinical stage T1-3N0- Medicine, Philadelphia/UNITED STATES OF AMERICA, 2Division Of 1M0 disease, PS 0-1, and no major comorbidity. In addition of routine Pulmonary, Allergy And Immunology, University Of Pennsylvania School Of preoperative chest CT, brain MRI, FDG-PET, and pulmonary V/Q scan, Medicine, Philadelphia/UNITED STATES OF AMERICA extended surgical staging was performed in 21 patients. Background: Immunotherapy approaches have improved endogenous anti- Results: A total of 61 patients (M/F 51/10, median age 62, range tumor immune responses in patients with minimal tumor burden; however, 37-71) were enrolled into EPP program after detailed evaluation by results are less dramatic in bulky tumor states. One potential explanation multidisciplinary team. Histology was epithelioid (n=53), biphasic (n=3), is that bulky tumors develop complex immunosuppressive networks. sarcomatoid (n=3), small cell mesothelioma (n=1), and unidentified (very Cytoreductive surgery has been shown to improve immunotherapy potency poster sessions | september 12 early, n=1). Clinical stage of 61 ITT patients and pathological stage for 45 in bulky tumor states; however, an exact mechanism remains unclear. We EPP patients were as follows: c-stage I/II/III 18/28/15, and p-stage I/II/ hypothesized that cytoreductive surgery reduces these immunosuppressive III/IV 3/10/28/4. Three patients underwent EPP without preoperative networks and restores the potency of immunotherapy in models of treatment, and they received either of chemotherapy or radiotherapy malignant mesothelioma. postoperatively. The remaining 58 received induction chemotherapy with cisplatin plus pemetrexed (n=49) or other regimens (n=9). Fifty-seven Methods: To test this hypothesis, a vaccine against the common out of the above 58 completed chemotherapy. In 9 patients, EPP was mesothelioma antigen, mesothelin, (Listeria.mesothelin) was tested in abandoned due to medical reasons (n=2) or patient’s refusal (n=7), and several syngeneic, murine models of mesothelioma. Cytoreductive surgery the remaining 48 started EPP. Only exploratory thoracotomy was given in was performed following treatment of advanced tumors. Mechanistic 6 patients due to unexpected extensive disease, and 42 completed EPP. underpinnings were investigated using flow cytometry, in vivo leukocyte In a total of 45 EPPs, macroscopic complete resection was achieved in 42 depletion methods, in vivo tumor neutralization assays and ELISAs. (69% of ITT). Of 50 candidates fortrimodality therapy, 26 (52%) completed all the treatment. All but one operation were performed by one surgeon Results: Mesothelioma vaccines were effective in treating small (SH). Surgical mortality rate at 30- and 90-days were 2.0% (1/51, bleeding, mesothelioma cancers, but had little anti-tumor effects in bulky cancer POD15) and 5.9% (ARDS, POD48 and 72). Surgical morbidity (CTCAE states. Interestingly, in bulky disease scenarios, surgical cytoreduction grade > 3) occurred in 9 patients (17.6%). The median, 2-year, and 5-year unmasked the anti-tumor potency of vaccine approaches. Immune survivals for ITT population (n=61) were 25.2 months, 52.3%, 28.1%, and mechanisms that explained restoration in anti-tumor immune responses 28.5months, 54.6%, 34.8% for patients who completed EPP (n=45), and included reduced myeloid derived suppressor cell populations, decreased 22.8 months, 46.0%, and 0% for no-EPP (n=16), respectively. immunosuppressive cytokine (IL-6, G-CSF, IL-10) levels, and augmented intratumoral CD8 T-cell trafficking.

Conclusion: This study provides the first report of a mechanism for the synergy between cytoreductive surgery and immunotherapy. This study also demonstrates that surgical resection combined with immunotherapy may be a rational therapeutic option for patients with malignant mesothelioma. international mesothelioma interest group

Disclosure: No significant relationships.

POSTER SESSION 1 September 12, 2012 11:30-12:30

P1.07: IS THERE ANY BENEFIT IN LUNG SPARING MACROSCOPIC COMPLETE RESECTION OVER VIDEO ASSISTED DEBULKING IN MALIGNANT PLEURAL MESOTHELIOMA?

David A. Waller, Apostolos Nakas Thoracic Surgery, Glenfield Hospital, Leicester/UNITED KINGDOM Conclusion: EPP is a highly invasive surgery but is feasible when performed by dedicated multidisciplinary team. Although better survival was observed Background: The role of lung sparing surgery to prolong survival in in no-EPP patients until 1.5 years after diagnosis, long-term survival was the management of Malignant Pleural Mesothelioma (MPM) is yet to be seen only in EPP patients. confirmed. Video Assisted Thoracoscopic Debulking Surgery (VATS PD) has a role in symptom control and may prolong survival. Our aim was to Disclosure: No significant relationships. determine whether Extended Pleurectomy Decortication (EPD) conveys any survival benefit over VATS PD.

Methods: From a prospective database we identified 224 patients (191 male) who underwent EPD (142) and VATS PD (82 patients) over 14 years. The patients in the EPD group were younger (mean 61.6 years) than the VATS group (mean 67.3, p<0.05). Cell type was similar ( 108 Epithelioid and 34 biphasic in the EPD group, 68 Epithelioid and 14 biphasic in the VATS group, p=0.24). We tested for differences in survival between the groups using selected subgroups.

iMig2012.org • Abstract Book 83 Results: Median follow up was 11 (1-90) months. 30 and 90 day mortality operative chemotherapy (CT); 41 (65%) received heated intra-operative was similar between the groups (4.9% EPD vs 4.8% VATS PD, p=1 and chemotherapy (HIOC); and 36 (59%) received adjuvant chemotherapy 10.6% vs 9.7 %, p=1 respectively). (usually cisplatin and/or pemetrexed). Median follow-up for the living From the data available approximately 50% had chemotherapy in both patients was 49 months (range 11-97). Among the 59 patients, 44 (75%) groups (p=0.57). developed recurrence and median time to recurrence was 11.1 months. Overall survival was similar between the groups: Median 14.4 (SE 1.7, 95% Forty-two patients (71%) have died. Median overall survival was 24.6 CI 11-18)months) for EPD compared to 13 (SE 1, 95% CI 11-15) months for months and 5-year overall survival was 27%. Five-year freedom from VATS, p=0.149. recurrence was 19%. Patterns of recurrence for all patients are shown in From the EPD group we identified a smaller subgroup of node negative the table. patients (n=45): The survival of this subgroup was slightly better ( median 16, SE 3.3, 95% CI 9-23 months), p=0.059 than for VATS. SITE OF RECURRENCE N % of All Patients % of Recurrences In patients with Epithelioid disease we observed marginally better survival in the EPD group ( n=108, median 18.7, SE 3, 95% CI 13-24 months) IHT* and/or 42 71 % 95 % compared to VATS PD (n=68, median 14.1, SE 1.2, 95% CI 12-16.4 months), Mediastinum p=0.079. Abdomen 9 15 % 20 % In contrast, in patients with Biphasic cell type, the extent of surgery had no effect on survival ( EPD n=34, median 10, SE 1.2, 95% CI 7.6-12 months Contralateral 7 12 % 16 % versus VATS PD n=14, median 8, SE 2.5, 95% CI 3-13 months, p=0.67). Hemithorax poster sessions | september 12 It was only the smaller subgroup of Epithelioid node negative EPD (n=28) Distant 1 2 % 2 % that demonstrated significantly better survival (median 27, SE 10, 97% CI 7.2-46.6 months) compared to Epithelioid VATS PD (n=68, median survival 14.1, SE 1.2, 95% CI 11.8-16.4 months), p=0.012. *IHT: ipsilateral hemithorax Twenty patients (45% of recurrences) failed in Conclusion: In most patients with mesothelioma, VATS debulking is all the ipsilateral hemithorax (IHT) only and 29 patients (66% of recurrences) that is indicated .Only node negative patients with Epithelioid disease failed in the ipsilateral hemithorax and/or mediastinum only. The most can benefit from the increased survival conveyed by EPD. This has many common specific sites of recurrence in the IHT were neo-pleural mass clinical implications including accurate histopathologic typing and invasive (71% of recurrences) and chest wall mass (36%); in the mediastinum preoperative staging of the disease to exclude nodal disease. were lymph nodes (100%); in the abdomen were abdominal mass (60%), The survival demonstrated in the Biphasic subgroup suggests that the retroperitoneal nodes (30%), and ascites (20%); in the CHT were lung behaviour of this type closely resembles the one demonstrated by nodules (57%) and pleural mass (29%); and the single distant recurrence Sarcomatoid MPM. was in bone.

Disclosure: No significant relationships. Conclusion: Most recurrences following pleurectomy for mesothelioma occur in the ipsilateral hemithorax. Future strategies should focus on methods to intensify local treatment such as HIOC, photodynamic therapy, systemic chemotherapy, radiotherapy or more aggressive resection POSTER SESSION 1 September 12, 2012 11:30-12:30 techniques.

P1.08: PATTERNS OF RECURRENCE FOLLOWING PLEURECTOMY/ Disclosure: No significant relationships. DECORTICATION FOR MALIGNANT PLEURAL MESOTHELIOMA

Andrea S. Wolf1, Ritu R. Gill2, Elizabeth H. Baldini3, Raymond H. Mak4, David E. Kozono4, Aileen B. Chen4, Marcelo Dasilva1, William G. Richards1, Raphael Bueno1, David J. Sugarbaker1 international mesothelioma interest group 1Division Of Thoracic Surgery, Brigham And Women’s Hospital And Harvard Medical School, Boston/MA/UNITED STATES OF AMERICA, 2Radiology, Brigham And Women’s Hospital, Boston/UNITED STATES OF AMERICA, 3Radiation Oncology, Brigham And Women’s Hospital/Dana-Farber Cancer Institute, Boston/MA/UNITED STATES OF AMERICA, 4Radiation Oncology, Brigham And Women’s Hospital/Dana- Farber Cancer Institute, MA/UNITED STATES OF AMERICA

Background: We sought to define patterns of recurrence following pleurectomy/decortication for patients with malignant pleural mesothelioma to inform future treatment strategies.

Methods: Under an IRB-approved protocol, we reviewed records for 59 consecutive patients with mesothelioma who underwent pleurectomy/ decortication without prior chemotherapy at our institution between 2001 and 2010. Pleurectomy was performed for macroscopic complete resection in patients with small volume tumor not involving the lung fissures or parenchyma and in those with contraindication to pneumonectomy. Data for treatment, recurrence and survival were retrieved from medical records. Surveillance CT or PET-CT scans were generally obtained every 3-6 months. A dedicated thoracic radiologist reviewed the post-operative chest CT scans and/or PET-CT scans to determine sites of recurrence. Time-to-recurrence and overall survival from date of pleurectomy were estimated by the Kaplan-Meier method.

Results: Median age was 69 years (range 27-86), and 44 patients (75%) were men. Median tumor volume was 73 cm3 (range 0 – 1814). Histology from pleurectomy specimens was epithelial for 45 patients (76%), and non-epithelial for 14 (24%). No patients received pre-

iMig2012.org • Abstract Book 84 Poster Session 1 Novel Therapeutics September 12, 2012 11:30-12:30

POSTER SESSION 1 September 12, 2012 11:30-12:30 POSTER SESSION 1 September 12, 2012 11:30-12:30

P1.09: THERMAL THERAPY IN THE TREATMENT OF MALIGNANT P1.10: PHASE 3 TRIAL OF NGR-HTNF PLUS BEST INVESTIGATOR’S PLEURAL MESOTHELIOMA CHOICE (BIC) VERSUS PLACEBO PLUS BIC IN PREVIOUSLY TREATED PATIENTS WITH MALIGNANT PLEURAL MESOTHELIOMA Robert B. Cameron, Dongmei Hou (MPM) poster sessions | september 12 Thoracic Surgery, David Geffen School Of Medicine At UCLA, Los Angeles/ CA/UNITED STATES OF AMERICA Vanesa Gregorc1, Gilda Rossoni1, Antonio Lambiase2, Claudio Bordignon2 1Oncology, Istituto Scientifico San Raffaele, Milan/ITALY, 2Clinical Background: Thermal therapy, both hyper- and hypothermia, has been Development, Molmed/ITALY used in cancer therapy for decades and hyperthermic chemotherapy perfusion, specifically, has been used in the treatment of mesothelioma Background: NGR-hTNF exploits the tumor-homing peptide asparagine- without data as to the optimal conditions. We sought to define in vitro the glycine-arginine (NGR) for selectively targeting tumor necrosis factor (TNF) most effective strategy for the use of thermal therapy in mesothelioma. to a CD13 overexpressed by cancer endothelial cells. MPM has one of the highest vascular endothelial growth factor levels among solid tumors and Methods: Growth of various in vitro established cell lines, including a tumor hypervascularity, as assessed by increased microvessel density, is hyperthermia-sensitive Chinese Hamster Ovarian cell line (CHO-K1), a independent predictor of poor overall survival (OS). In a single-agent phase normal lung fibroblast line (MRC-5), a lung cancer line (A549), and three 2 trial (JCO 28:2604,2010) on 57 pemetrexed-pretreated MPM patients human mesothelioma cell lines (NCI-H28, NCI-H2052, and MSTO-211H) (pts), a disease control (DC) rate of 46% lasting for a median time of 4.7 exposed to varying hyper- and hypothermic conditions was investigated months (95% CI 4.0-5.4), and a median OS of 12.1 months (7.2-17.0) using either a standard metabolic MTS absorbance assay at 490 nm or were reported. Notably, a DC rate of 50% lasting for a median time of a standard clonogenic assay, enumerating colony-forming units >50 9.1 months (4.7-13.4) was noted among a small cohort receiving weekly cells. Each cell line was expanded in flasks and then exposed to varying dosing. combinations of hyperthermia (37-45o C), hypothermia (0-[-80]o C), and/or chemotherapy. Cells were harvested and assays performed. Methods: MPM pts who are failing a pemetrexed-based chemotherapy Chemotherapeutic agents used were cisplatin, gemicitabine, and are currently enrolled in a 2-arm (1:1 ratio), placebo-controlled phase 3 pemetrexed. trial. BIC includes either supportive care alone or combined with single- agent chemotherapy (doxorubicin, gemcitabine, or vinorelbine). Before Results: Initially, a dose-response curve was generated using hyperthermia randomization, the physician has to decide for each patient if he/she is o o alone in CHO-K1, A549, and NCI-H28 cell lines heated to 37 , 42 , and candidate to either supportive care alone or combined with chemotherapy. o 45 for 20, 40, and 60 mins. This showed a reproducible dose-response Pts are randomly assigned to the treatment group by the following o effect in CHO-K1 cells reaching a survival nadir of 1.5% of controls at 45 for stratification factors: candidate for chemotherapy (yes vs no), type of international mesothelioma interest group 60’ (p<0.01). A549 cells and NCI-H28 mesothelioma cells showed similar chemotherapy, and ECOG performance status (0 vs 1-2). less dramatic responses however NCI-H28 mesothelioma cell growth o was only reduced a modest 65% at 45 for 60’ (p<0.01). Comparison of Results: NGR-hTNF/placebo is given intravenously as 1-h infusion at 0.8 two assays showed that measurement of metabolic activity (MTS assay) µg/m2 weekly until progressive disease. Best supportive care is delivered failed to demonstrate the differences documented by the clonogenic according to institutional and literature guidelines. Primary study endpoint assay and therefore was the MTS assay was abandoned. The addition of is OS, while secondary endpoints include progression free survival, DC rate chemotherapy to hyperthermia was then assessed. Doses were chosen (with CT-scan repeated every 6 weeks according to MPM-modified RECIST based on prior pharmacokinetic data from studies showing a maximum criteria), safety, and quality of life. A correlative study for pharmacokinetic tissue/blood level of 200 ngm/ml for cisplatin pleural instillation and were as well as for plasma/tumor biomarkers assessment is included. For felt to more accurately reflect actual tumor tissue levels. In these studies, primary analysis of OS duration, 390 patients are required to detect a o the clinically-relevant limit of 42 was used for hyperthermia. Cisplatin hazard ratio of 0.72 with at least 80% power for a 2-sided 0.05-level test. alone reduced the clonogenic potential modestly to 26%, 16.4%, and 13.6% at 42o for 60’ (p<0.01); however, this was only a further reduction of Conclusion: The study (NGR015) is currently open to accrual in EU, US, 29.6%, 33.8%, and 34.2%, respectively, from the cisplatin alone control. Canada, and Egypt (ClinicalTrials.goVNCT01098266) with more than half of Therefore, most of the reduction was attributable to chemotherapy not patients recruited so far. hyperthermia. With combinations of cisplatin/gemcitabine and cisplatin/ pemetrexed the effect was greater, with reduction to 9.6%, 0%, and 0% Disclosure: Antonio Lambiase -Employment - MolMed Claudio Bordignon (p<0.01) (incremental reduction of only 16.5%, 0%, and 0%, respectively - Employment - MolMed from hyperthermia). Cisplatin/pemetexed produced essentially identical results. The effect of hypothermia alone was then assessed. Exposure to freezing temperature (-80oC) reduced the clonogenic potential of all cells lines, including the 3 mesothelioma lines, in a dose-dependent manner by POSTER SESSION 1 September 12, 2012 11:30-12:30 >99% in as little as 5 minutes (p<0.01). Further, no particular sensitivity was noted for mesothelioma cell lines (compared to other cells) to hyper- P1.11: REPLICATION-COMPETENT RETROVIRUS VECTOR- or hypothermia or even chemotherapy. MEDIATED SUICIDE GENE THERAPY IN EXPERIMENTAL MODELS OF HUMAN MALIGNANT MESOTHELIOMA Conclusion: Chemotherapy appears to be most effective when using two drug combinations. Thermal therapy appears to be most effective when Shuji Kubo1, Misato Takagi-Kimura1, Atsuko Tamamoto1, Tomoko using hypothermia rather than hyperthermia and this modality warrants Hashimoto-Tamaoki1, Noriyuki Kasahara2 further investigation.

Disclosure: No significant relationships. iMig2012.org • Abstract Book 85 1Genetics, Hyogo College Of Medicine, Nishinomiya/JAPAN, 2Medicine between 93 patients with High or Low ERβ expressing MMe tissue. Among And Molecular & Medical Pharmacology, UCLA David Geffen School Of genes down-regulated in the High ERβ group we identified the sdhb gene, Medicine, Los Angeles/CA/UNITED STATES OF AMERICA coding for the mitochondrial respiratory chain (MRC) complex II SDHB subunit, and among the up-regulated the KDM6B gene, coding for the Background: Replication-competent retrovirus (RCR) vectors have been α-ketoglutarate dependent histone H3 Lys-27 demethylase. The aim of shown to achieve significantly enhanced tumor transduction efficiency and our work was therefore to evaluate the relationships between these genes therapeutic efficacy in various cancer models. In the present study, we and assess whether ERβ acts as a tumor suppressor by interfering with investigated RCR vector-mediated suicide gene therapy for the treatment mitochondrial oxidative energy metabolism. of malignant mesothelioma, a highly aggressive tumor with poor prognosis. Results: In in vitro cell models, we demonstrated that sdhb gene down- Methods: To evaluate the utility and efficiency of RCR vectors for gene regulation induced a significant increase in ERβ gene expression, in a delivery to mesothelioma cells, RCR-GFP vector, expressing the green KDMB6 dependent manner. ERβ activation with the specific agonist fluorescent protein (GFP) marker gene, was first tested on a panel of human KB9520, significantly compromised both MRC complexes activity and malignant mesothelioma and non-malignant transformed mesothelial cell mitochondrial ATP production. These data were confirmed in vivo in lines in vitro. Transduction efficiency and replicative spread of the RCR MMe mice models. In addition, ERβ over-expression caused a severe vector over time was monitored by flow cytometry and in vivo imaging. mitochondrial impairment and led to an increased glucose-dependence. Next, to evaluate the potential of RCR vector-mediated suicide gene

therapy for this malignancy, we employed RCR-yCD, expressing the yeast Conclusion: On the whole our data suggest that selective targeting of ERβ poster sessions | september 12 cytosine deaminase (yCD) suicide gene. Following administration of the may be an efficacious treatment option of this neoplasm. prodrug 5-fluorocytosine (5FC), cytotoxicity against RCR-yCD infected malignant mesothelioma cells was assessed in vitro by Alamar blue Disclosure: No significant relationships. assay, and tumor growth inhibition effects in vivo were assessed in both subcutaneous xenograft tumor and disseminated peritoneal tumor models of malignant mesothelioma. POSTER SESSION 1 September 12, 2012 11:30-12:30 Results: RCR-GFP vector successfully infected and efficiently replicated in human malignant mesothelioma cell lines, as compared with non- P1.13: EFFICACY AND TOXICITY OBSERVED IN MALIGNANT malignant mesothelial cells in vitro. In mice with pre-established PLEURAL MESOTHELIOMA PATIENTS TREATED IN PHASE I TRIALS subcutaneous tumor xenografts, RCR-GFP vector showed robust spread AT A SINGLE INSTITUTION throughout entire tumor masses after intratumoral administration. Next, RCR-CD, expressing the yeast CD suicide gene, showed efficient Jacques Raphael1, Antoine Hollebecque2, Gwénaël Le Teuff3, Christophe transmission of the suicide gene associated with replicative spread of the Massard2, Rastislav Bahleda2, Jacques Margery4, Benjamin Besse1, Jean- virus, resulting in efficient killing of malignant mesothelioma cells in a charles Soria1, David Planchard1 prodrug 5FC-dose dependent manner in vitro. After a single intratumoral 1Thoracic Group, Inserm U981, Institut Gustave Roussy, Villejuif/ injection of RCR-CD followed by intraperitoneal administration of 5FC, FRANCE, 2Medical Oncology Department, Institut Gustave Roussy, Villejuif/ RCR vector-mediated suicide gene therapy achieved significant inhibition FRANCE, 3Department Of Statistics And Epidemiology, Institut Gustave of subcutaneous tumor growth, and significantly prolonged survival in the Roussy, Villejuif/FRANCE, 4Pulmonary Department, Percy Hospital, Paris/ disseminated peritoneal model of malignant mesothelioma. FRANCE

Conclusion: These data indicate the potential utility of RCR vector- Background: Malignant Pleural Mesothelioma (MPM) is a locally mediated suicide gene therapy in the treatment of malignant aggressive disease with a poor prognosis. After failure of platinum-based mesothelioma. chemotherapy in first line, there is no widely approved salvage regimen. New strategies for treatment of MPM are needed and phase I trials appear Disclosure: No significant relationships. as a rationale alternative. The results of such an approach have been international mesothelioma interest group evaluated.

Methods: MPM patients, were enrolled in 20 different phase I trials POSTER SESSION 1 September 12, 2012 11:30-12:30 between March 2005 and January 2012, and their data analysed retrospectively. The primary endpoint was response rate and secondary P1.12: ESTROGEN RECEPTOR β ACTIVATION IMPAIRS endpoints were toxicity profile, Overall Survival (OS) and Progression Free MITOCHONDRIAL OXIDATIVE ENERGY METABOLISM AND Survival (PFS). OS was defined as the time from the date of inclusion in AFFECTS MALIGNANT MESOTHELIOMA CELL PROLIFERATION IN a phase I till the death or the date of last news. PFS was defined as the VITRO AND IN VIVO time from the date of inclusion till the radiological (or clinical) progression if it occurs within the phase I otherwise the date of withdrawal. Median Arcangela Gabriella Manente1, Daniela Valenti2, Giulia Pinton3, Puthen follow-up of patients was estimated through Schemper’s method. The cut- Jithesh4, Dean Fennell5, Antonio Daga6, Leonardo Rossi7, Steven off date for the analysis was April 2012. Adverse events were assessed by Gray8, Kenneth O’Byrne8, Anna Rosa Vacca2, Stefan Nilsson9, Luciano NCI-CTC v.3.0 and response rate according to RECIST 1.1 criteria (CT-scan Mutti10, Laura Moro3 every 4-6 weeks). 1Dept. Of Pharmacological Sciences, University Of Piemonte Orientale A. Avogadro, Novara/ITALY, 2Cnr-Ibbe, Bari/ITALY, 3Dept. Of Pharmaceutical Results: Forty-six patients with a confirmed histology of MPM were Sciences, University Of Piemonte Orientale “A. Avogadro”, Novara/ included in the analysis with a median follow up of 20 months. The ITALY, 4University Of Liverpool, Liverpool/UNITED KINGDOM, 5Mrc median age was 61 years old, sex ratio (M/F) was 2.29 and histological Toxicology Unit, University Of Leicester, Leicester/UNITED KINGDOM, 6Irccs type was mainly epitheloid (78%). Eastern Cooperative Oncology Group San Martino-Ist, Genova/ITALY, 7University Of Pisa, Pisa/ITALY, 8Institute performance status was <2 in 96% of pts. Sixty-three percent of pts Of Molecular Medicine, St James’s Hospital, Dublin/IRELAND, 9Karo Bio Ab, received more than one line of platinum-based chemotherapy before the Huddinge/SWEDEN, 10Vercelli Hospital, Vercelli/ITALY phase I trial. Radiological disease progression was observed in 50% of pts, clinical progression in 28%, and dose limiting toxicity in 22%. The best Background: Estrogen receptor (ER) β has been already shown to act as tumour response was as follows: 7% of pts had a RECIST partial response, a tumor suppressive gene in Malignant Mesothelioma (MMe) and to be a 59% had stable disease for a median duration of 2.8 months and 24% had prognostic factor of longer patient survival. progressive disease. The median treatment duration in the phase I trial was 1.9 months. Median OS and PFS were 6 months (95% CI= [4.3-10.7]) and Methods: By in silico analysis of microarray data we generated an ERβ gene 2.1 months (95% CI= [1.3-2.8]) respectively. The most common grade 3/4 expression meta-signature, identifying 172 genes differentially expressed adverse events (41%) were haematological (11%), gastro-intestinal (4%),

iMig2012.org • Abstract Book 86 cutaneous (7%), renal (4%) and hepatic (4%). All adverse events were POSTER SESSION 1 September 12, 2012 11:30-12:30 reversible and no death due to therapy toxicity was reported. P1.15: SWITCH MAINTENANCE TREATMENT WITH NGR-HTNF OR Conclusion: These results show that including MPM pts in phase I PLACEBO AFTER COMPLETION OF FRONT-LINE CHEMOTHERAPY trials beyond first line of treatment can result in clinical benefits with IN PATIENTS WITH MALIGNANT PLEURAL MESOTHELIOMA (MPM) an acceptable toxicity profile. Several molecular pathways involved in MPM have been identified and further novel biologic therapies might be Paolo Zucali1, Manlio Mencoboni2, Federica Grosso3, Camillo Porta4, tested in a phase I setting in a biology-oriented approach rather than a Armando Santoro1, Antonio Lambiase5, Claudio Bordignon5 stochastical one. We are currently offering a tumour molecular profiling in 1Department Of Oncology, Humanitas Cancer Center Irccs, Rozzano/ our pts (MOSCATO trial) for a better selection of phase I trial. ITALY, 2Medical Oncology, Villa Scassi Hospital Asl 3 Genovese, Genova/ ITALY, 3Oncologia, Ospedale Civile, Alessandria/ITALY, 4Dipartimento Di Disclosure: No significant relationships. Oncologia, Ospedale, Pavia/ITALY, 5Clinical Development, Molmed/ITALY

Background: NGR-hTNF exploits the asparagine-glycine-arginine (NGR) peptide for selectively targeting tumor necrosis factor (TNF) to a CD13 POSTER SESSION 1 September 12, 2012 11:30-12:30 overexpressed on cancer endothelial cells. Tumor hypervascularity is an independent predictor of poor overall survival (OS) in MPM. In a phase P1.14: FAK INHIBITOR VS-4718 PREFERENTIALLY ATTENUATES 2 trial on 57 MPM patients (pts) with progressive disease (PD) after a poster sessions | september 12 CELL GROWTH OF MALIGNANT MESOTHELIOMAS WITH NF2 pemetrexed-based regimen (JCO 2010, 2604-11), we reported a disease MUTATION: ROLE OF CANCER STEM CELLS control rate of 46% (26/57 pts; 95% CI 32-59), maintained for a median time of 4.7 months (4.0-5.4), and a median OS of 12.1 months (7.2-17.0). Irina M. Shapiro1, Vihren N. Kolev1, Christian M. Vidal1, Mitchell Keegan1, Recent data in lung cancer pts have shown that a maintenance treatment Qunli Xu1, Craig Menges2, Joseph Testa2, Jonathan A. Pachter1 given immediately after standard 1st line therapy can improve PFS and 1Suite 440, Verastem, Inc., Cambridge/UNITED STATES OF OS (JCO 2011, 3825-31). Standard 1st line in advanced MPM is based on AMERICA, 2Prevention Pavillion Room 3040, Fox Chase Cancer Center, 6 cycles of a pemetrexed-based therapy, with a median progression free Philadelphia/UNITED STATES OF AMERICA survival (PFS) of about 6 months. However, the median time from 1st line completion to 2nd line initiation is only 3 months (Ann Oncol 2005, 923- Background: Malignant pleural mesothelioma (MPM) is an aggressive 927). tumor in the pleural lining of the lung often caused by asbestos exposure. MPM patients are usually diagnosed at an advanced stage of the disease Methods: This is a multicenter, double-blind, 2-arm (1:1 ratio), randomized and the prognosis is poor. Median survival after diagnosis is 9 to 12 phase 2 trial with a comparison of NGR-hTNF vs placebo in advanced MPM months and standard-of-care agents such as cisplatin and pemetrexed are patients who did not progress after 6 cycles of a front-line, pemetrexed- relatively ineffective in increasing median survival time for MPM patients. based regimen. Maintenance treatment is started within 3 to 7 weeks from New therapeutic modalities are urgently needed to improve the prognosis the last chemotherapy cycle, including 3 weeks of wash-out post-therapy of MPM patients. Neurofibromatosis 2 (NF2) is a tumor suppressor gene and 4 weeks for assessment. Patients are randomly assigned to treatment that encodes the protein Merlin. Biallelic inactivation of NF2 by mutation groups using ECOG performance status (0 or 1) as stratification factor. and/or deletion occurs in ~40% of MPMs leading to inactive Merlin. Merlin has been demonstrated to play roles in cell adhesion, invasion Results: NGR-hTNF/placebo is given intravenously as 1 hour infusion and cell motility in tumor cell lines partially through regulation of focal at 0.8 µg/m2 weekly until PD. Primary endpoint is PFS computed from adhesion kinase (FAK) which in turn mediates signal transduction by randomization date, with patient unblinding allowed after PD. Secondary integrins and growth factor receptors. Increased activation of FAK has been endpoints include OS, tumor response (assessed every 6 weeks by MPM- demonstrated in NF2-mutated mesothelioma cells, indicating that FAK may modified RECIST criteria), safety, and quality of life. A correlative study represent an important therapeutic target for MPM. for testing plasma/tumor biomarkers is included. The study is designed to have 80% power to reject the null hypothesis of no difference at the international mesothelioma interest group Methods: Using 3D cell growth analysis, we tested effects of the potent 0.1 significance level (1-sided) if the experimental arm is associated with a and selective FAK inhibitor VS-4718 on cell growth and viability in a panel reduction in the hazard of progression or death of 38% (HR= 0.62). of mesothelioma cell lines. An ALDEFLUOR assay was used to assess the percentage of cancer stem cells (CSCs) in MPM cell lines treated with Conclusion: Seventeen patients have been enrolled so far. ClinicalTrials.gov VS-4718 in comparison to the standard-of-care agents pemetrexed and NCT01358084. cisplatin. Disclosure: Antonio Lambiase - Employment - MolMed Claudio Bordignon Results: VS-4718 was evaluated in a panel of MPM cell lines with wild- - Employment - MolMed type or mutated NF2. Mutant NF2 MPM cells were found to be especially sensitive to the FAK inhibitor VS-4718 with EC50 values below 100 nM, in contrast to wild type NF2 MPM cell lines which were less sensitive with

EC50 values above 1 mM. Ectopic expression of a non-phosphorylatable artificial mutant of NF2 (NF2-S518A) in NF2 mutant MPM cells abolished the enhanced sensitivity to VS-4718, confirming the hypothesis that Merlin loss mediates sensitivity to VS-4718. Interestingly, MPM cell lines were found to have sub-populations of ALDEFLUOR+ CSCs. Furthermore, the FAK inhibitor VS-4718 induced a significant reduction in the percentage of CSCs in contrast to the standard-of-care agent pemetrexed which enriched the CSC population.

Conclusion: In summary, our results indicate that the FAK inhibitor VS-4718 is especially potent in NF2-mutated MPM tumor cells, and that NF2 status may be a valuable stratification marker for VS-4718 response. Furthermore, cancer stem cells in NF2 mutant mesothelioma appear to be particularly resistant to pemetrexed, but sensitive to VS-4718. We believe that these data support the clinical development of the selective FAK inhibitor VS-4718 for treatment of NF2-mutated malignant mesothelioma.

Disclosure: I am an employee and a stockholder of Verastem, Inc.

iMig2012.org • Abstract Book 87 Poster Session 1 Imaging and Staging September 12, 2012 11:30-12:30

POSTER SESSION 1 September 12, 2012 11:30-12:30 Conclusion: Analysis of measurement trajectories of lung volumes and disease volumes during chemotherapy for patients with mesothelioma P1.16: LUNG AND DISEASE VOLUME MEASUREMENTS AS indicates that decreasing lung volume and increasing disease volume are MARKERS FOR PATIENT RESPONSE IN MALIGNANT PLEURAL significantly and independently associated with poor patient prognosis. MESOTHELIOMA

Disclosure: No significant relationships. poster sessions | september 12 Zacariah E. Labby1, Anna K. Nowak2, H.L. Kindler3, Samuel G. Armato1 1Department Of Radiology, The University Of Chicago, Chicago/IL/UNITED STATES OF AMERICA, 2School Of Medicine And Pharmacology, University Of POSTER SESSION 1 September 12, 2012 11:30-12:30 Western Australia, Perth/WA/AUSTRALIA, 3Department Of Medicine, The University Of Chicago, Chicago/IL/UNITED STATES OF AMERICA P1.17: OBSERVER VARIABILITY IN MESOTHELIOMA TUMOR Background: The current standard for image-based response assessment THICKNESS MEASUREMENTS in mesothelioma is the Modified RECIST measurement technique with 1 2 2 changes classified according to the standard RECIST criteria. The purpose Samuel G. Armato , Roslyn Francis , Anna K. Nowak 1 of this study was to investigate continuous changes in volumetric Department Of Radiology, The University Of Chicago, Chicago/IL/UNITED 2 measurements during treatment as a marker of response for mesothelioma STATES OF AMERICA, School Of Medicine And Pharmacology, University Of patients. Both disease volumes and lung volumes, a physiological correlate Western Australia, Perth/AUSTRALIA of disease volumes, were investigated in this study. Background: Single time-point unidimensional tumor thickness Methods: Serial follow-up CT scans were retrospectively obtained during measurements define measurable disease for clinical trial inclusion and also the course of clinically standard chemotherapy for 61 patients in this IRB- constitute a field in the IASLC prospective mesothelioma staging database. approved study. For each of the 216 CT scans the aerated lung volumes It is unclear how tumor thickness, morphology, and location affect were segmented using a fully automated method, and the pleural disease interobserver variability in a single baseline measurement. volume was segmented using a semi-automated method. Diseased (ipsilateral) lung volumes were normalized by the respective contralateral Methods: 105 thoracic CT scans were collected retrospectively from 50 lung volumes to account for differences in respiratory phase between the mesothelioma patients. Each scan was reviewed by a medical oncologist, scans of each patient. Relative changes in each measurement technique who identified 170 sites of mesothelioma tumor across all scans that from baseline were tracked over the course of serial follow-up imaging. represented a range of thickness, lesion morphology (concave rind, Survival modeling was performed using time-varying Cox proportional convex rind, convex mass, fusiform mass), and location. Using a custom hazards models. computer interface, reference tumor thickness measurements were

obtained by creating a line segment that spanned the tumor from the international mesothelioma interest group Results: Median survival from baseline imaging prior to treatment initiation outer tumor margin along the chest wall or mediastinal structures to the was 12.7 months (95% confidence interval, 10.2–15.3 months). Over the inner tumor margin. An observer study was conducted in which each of course of treatment, disease volume decreased by an average of 19% from five other physicians was presented with the individual CT sections and baseline, and normalized lung volume increased an average of 8% from the same fixed location of the outer tumor margin at each of the 170 baseline. When discretized as lung volume gain versus lung volume loss pre-defined tumor measurement sites. Each observer then independently over the course of treatment, patients experiencing lung volume loss (n=25) created a line segment to capture tumor thickness at all measurement had significantly worse prognosis than patients with lung volume gain sites. Interobserver variability was calculated as a function of tumor (n=36; p=0.0003). A strong negative correlation was observed between reference thickness to identify the smallest tumor thickness at which linear measurements of lung volume and disease volume, and as continuous measurements could be made reliably. Comparisons were made with the numerical parameters, both normalized lung volume and disease volume RECIST tumor response criterion of 20% for progression. were significant imaging biomarkers of patient prognosis in independent survival models. Results: Measurements acquired at tumor sites with reference thickness less than 7.5 mm demonstrated inter-observer variability (as defined by the difference between the maximum and minimum measurements of the observers at each site then tabulated across all sites in a specific size range) with a 75th percentile that included 20% of the tumor length. Inter-observer variability did not differ across lesion morphologies and locations. Only tumor sites with reference thickness greater than 12.5 mm demonstrated inter-observer variability with a maximum value that did not include 20% of the tumor length.

Conclusion: The results of this study have implications for the definition of minimum measurable tumor adopted by clinical trial and staging protocols.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 88 POSTER SESSION 1 September 12, 2012 11:30-12:30 Methods: 26 patients underwent radical pleurectomy with intraoperative photodynamic therapy as part of a surgery-based treatment for malignant P1.18: IMPORTANCE OF CERVICAL MEDIASTINOSCOPY AND pleural mesothelioma. Each specimen was removed and placed in a graded LAPAROSCOPY IN DEFINING TREATMENT OF MALIGNANT vessel containing saline, whereby its volume was determined by simple PLEURAL MESOTHELIOMA displacement.

Evelien De Witte1, Philippe Nafteux1, Kristiaan Nackaerts2, Eric Results: The stage breakdown for all 26 patients was: 2 Stage I, 1 Stage II, Verbeken3, Marc Decramer2, Karin Haustermans4, Paul De Leyn1 20 Stage III, and 3 Stage IV. 21 patients had an epithelial subtype and 5 had 1Thoracic Surgery, University Hospital, Leuven/BELGIUM, 2Pneumonology, a nonepithelial subtype. Overall, these patients had a median interquartile University Hospital Leuven, Leuven/BELGIUM, 3Pathology, University tumor volume of 625 (325, 888) cc. Patients with nonepithelial subtypes Hospital Leuven, Leuven/BELGIUM, 4Radiotherapy - Oncology, University had a significantly larger median tumor volume of 800 (800, 1500) cc, as Hospital Leuven, Leuven/BELGIUM compared to the median tumor volume of 450 (300, 850) cc in patients with the epithelial subtype (p=.047). Survival calculations were limited to Background: Multimodality treatment is an option in early stage malignant the 21 patients with the epithelial subtype (2 Stage I, 1 Stage II, 15 Stage III, pleural mesothelioma (MPM) but morbidity remains high. Therefore, 3 Stage IV) and were made at a mean/median follow up of 9.1/8.0 months. careful oncologic pre-treatment evaluation is mandatory. Standard Within this group, the median (95% CI) progression free survival (PFS) was clinical staging based on computed tomography, positron emission 38.3 (17.0, 54.0) months; median overall survival could not be calculated tomography and magnetic resonance imaging is often inaccurate. AIM: due to insufficient follow-up. Tumor volume was not associated with either poster sessions | september 12 To evaluate the added value of surgical staging, consisting of cervical PFS or overall survival. videomediastinoscopy (CM) and laparoscopy. Conclusion: Bearing in mind the limitations of this small retrospective Methods: Between March 2003 and December 2010, 126 patients with study, it appears that patients with nonepithelial tumors who enrolled for MPM were prospectively studied. Multimodality treatment was proposed surgery typically had greater tumor bulk. Focusing on the patients with in 82 fit patients based on standard clinic staging methods without epithelial tumors, for whom our group is currently limiting all surgery- taking into account presence of suspect mediastinal lymph nodes or based treatments, tumor volume did not appear to be a prognostic factor. transdiaphragmatic invasion. All patients underwent subsequent surgical At this time we conclude that this data will warrant a more mature analysis, staging (CM and laparoscopy (n=72), single CM (n=8) or single laparoscopy but there does not appear to be a reason to use tumor volume as an (n=2)). exclusion criteria for surgery-based treatment, at least for this particular therapeutic regimen. Given the ease with which tumor volume can be Results: Cervical videomediastinoscopy (n=80) caused node (N) determined for a radical pleurectomy specimen, we have now incorporated downstaging in 3 patients (3.75%), upstaging in 9 patients (11.25%) this measurement as part of every procedure. Furthermore, this technique and change from single level to multilevel N2 in 4 patients (5.00%). would also allow for very accurate calibrations by any radiographic Laparoscopy (n=74) discovered transdiaphragmatic invasion in 2.70% (n=2) algorithms designed to measure pleural tumor volume. and peritoneal metastasis in 2.70% (n=2) of patients. In 3 patients (4.05%) a suspicion of transdiaphragmatic invasion could not be confirmed during Disclosure: No significant relationships. laparoscopy. Surgical staging caused a change in treatment in 23.17% (n=19). Five patients became candidates for multimodality treatment (6.10%) and 14 patients were referred for non-surgical based treatment (17.07%).

Conclusion: Based on these results, we recommend that surgical staging consistent of CM and laparoscopy should be included in the standard evaluation of patients potentially candidates for multimodality treatment of MPM. international mesothelioma interest group

Disclosure: No significant relationships.

POSTER SESSION 1 September 12, 2012 11:30-12:30

P1.19: TUMOR VOLUME AS PROGNOSTIC FACTOR FOR PATIENTS UNDERGOING RADICAL PLEURECTOMY FOR MALIGNANT PLEURAL MESOTHELIOMA

Joseph Friedberg1, Melissa J. Culligan1, Mary Putt2, Benjamin French2, Stephen M. Hahn3, Charles B. Simone3, Evan Alley4, James Stevenson5, Daniel Sterman4, Eric J. Wang6, Keith Cengel3 1Thoracic Surgery, University Of Pennsylvania, Philadelphia/PA/ UNITED STATES OF AMERICA, 2Biostatistics And Epidemiology, University Of Pennsylvania, Philadelphia/PA/UNITED STATES OF AMERICA, 3Radiation Oncology, University Of Pennsylvania/UNITED STATES OF AMERICA, 4Medicine, University Of Pennsylvania, Philadelphia/ UNITED STATES OF AMERICA, 5Taussig Cancer Institute, Cleveland Clinic Foundation, Cleveland/OH/UNITED STATES OF AMERICA, 6Medical College Of Georgia/UNITED STATES OF AMERICA

Background: Our group performs radical pleurectomies to achieve a macroscopic complete resection in patients undergoing surgery-based therapy for malignant pleural mesothelioma. As a result, the surgical specimen is essentially all cancer, thereby allowing a very accurate determination of tumor volume. The purpose of this study is to determine whether tumor volume has prognostic value.

iMig2012.org • Abstract Book 89 POSTER SESSION 1 September 12, 2012 11:30-12:30 Ritu R. Gill1, William G. Richards2, Beow Y. Yeap3, Andrea S. Wolf2, Marcelo Dasilva2, Rapahel Bueno2, Elizabeth H. Baldini4, David J. P1.20: POSTERIOR INTERCOSTAL LYMPH NODES AND Sugarbaker5 MALIGNANT PLEURAL MESOTHELIOMA - WHAT ARE THEY, 1Radiology, Brigham And Women’s Hospital, Boston/MA/UNITED 2 WHERE ARE THEY AND WHAT DO THEY MEAN? STATES OF AMERICA, Division Of Thoracic Surgery, Brigham And Women’s Hospital And Harvard Medical School, Boston/MA/UNITED 3 Joseph Friedberg1, Melissa J. Culligan1, Mary Putt2, Benjamin French2, STATES OF AMERICA, Department Of Medicine, Massachusetts General Stephen M. Hahn3, Charles B. Simone3, James Stevenson4, Evan Alley5, Hospital And Harvard Medical School, Boston/MA/UNITED STATES OF 4 Daniel Sterman5, Eric J. Wang6, Keith Cengel3 AMERICA, Radiation Oncology, Brigham And Women’s Hospital/Dana- 5 1Thoracic Surgery, University Of Pennsylvania, Philadelphia/PA/UNITED Farber Cancer Institute, Boston/MA/UNITED STATES OF AMERICA, Division STATES OF AMERICA, 2Biostatistics And Epidemiology, University Of Of Thoracic Surgery, Brigham And Women’s Hospital, Boston/MA/UNITED Pennsylvania, Philadelphia/PA/UNITED STATES OF AMERICA, 3Radiation STATES OF AMERICA Oncology, University Of Pennsylvania/UNITED STATES OF AMERICA, 4Taussig Cancer Institute, Cleveland Clinic Foundation/UNITED Background: At our institution, indications for pleurectomy among STATES OF AMERICA, 5Medicine, University Of Pennsylvania, Philadelphia/ patients with MPM include patients with small volume disease and patients UNITED STATES OF AMERICA, 6Medical College Of Georgia/UNITED STATES who would require EPP for macroscopic complete resection but who have OF AMERICA other contraindications for pneumonectomy. poster sessions | september 12

Background: Little is known about the significance of the posterior Methods: Retrospective review of 59 patients with mesothelioma who intercostal lymph nodes, which are located within the intercostal spaces at underwent pleurectomy without prior chemotherapy at Brigham and the level of the rib heads. These nodes are not part of any staging system. Women’s Hospital between 2001 and 2010 was performed. Demographic, This report is an initial attempt to determine the significance of these clinical, pathologic, treatment, and survival data were collected. A nodes in patients with malignant pleural mesothelioma. dedicated thoracic radiologist reviewed CT and PET-CT scans to assess pre-operative disease distribution and post-operative recurrence. Tumor Methods: Posterior intercostal lymph nodes were harvested from 38 volume was calculated from pre-operative CT scans using Vitrea Enterprise patients with epithelial mesothelioma who underwent radical pleurectomy suite 6.0 (Vital Images, Minnesota, USA). Time to recurrence (TTR) and and intraoperative photodynamic therapy for malignant pleural overall survival (OS) were calculated from date of resection and estimated mesothelioma. The nodes were accessed by dividing the endothoracic by the Kaplan-Meier method. Cox regression models were derived using fascia at the level of the rib heads and bluntly dissecting into the intercostal significant univariate predictors at the 0.05 level. spaces. Results: Median age was 69 years (range 27-86). Forty-four patients Results: 18 out of 38 patients had positive intercostal nodes. Standard (75%) were men and 15 were women (25%). Histology determined by nodal status breakdown was 4/38 N0, 2/38 N1, and 32/38 N2. Positive biopsy was epithelial for 51 patients (86%) and non-epithelial for 8 (14%). 3 intercostal nodes were not associated with stage, but approached Median tumor volume was 73 cm (range 0 – 1814), and 37 patients significance with N status (Fisher’s Exact Test, p=0.073). At a mean/median (63%) had disease present in the fissures. No patients received pre- follow-up of 29.6/6.07 months, the group as a whole displayed a median operative chemotherapy (CT); 41 (65%) received heated intra-operative (95% CI) progression free survival of 12.3 (9.7- 20.1) months and overall chemotherapy (HIOC); and 35 (59%) received adjuvant CT. Forty-four survival of 24.6 (18.2, ND) months. There were no significant differences patients developed recurrent disease (75%) and 42 patients died (71%). between either progression free survival or overall survival among 30 Median TTR for all patients was 11.1 months. Median OS was 24.6 months, Stage III or 8 Stage IV patients. Patients with negative intercostal nodes and 5-year OS was 27%. Univariate analyses of predictors for TTR and OS had a median progression free survival of 16.4 (9.8, ND) months compared are listed in table. to 10.2 (9.5, 24.1) months for patients with negative intercostal nodes, but these differences did not achieve statistical significance (Log-rank

N Median p-value Median p-value international mesothelioma interest group test, p=.148). Patients with negative intercostal nodes demonstrated a TTR OS significantly longer median survival of 45.1 (19.4, ND) months compared (months) (months) to 16.8 (11.3, ND) months for patients with positive intercostal nodes (Log-rank test, p=.030). In a Cox model that included both stage and Age ≤ 69 Age 30 29 14.7 7.8 0.03 42.8 13.5 0.0005 intercostal nodes, positive intercostal nodes continued to be associated > 69 with increased risk of death (HR=3.02 (1.06, 8.65), p=.039). Statistical Female Male 15 44 11.7 10.5 0.8 42.8 20.8 0.07 evaluation of N status as a prognostic factor for progression free survival or Epithelial 51 8 11.7 10.5 0.4 27.6 15.3 0.2 death was not possible in these data due to the small number of subjects histology# Non- who were N0 or N1. epithelial histology Conclusion: Bearing in mind the limitations of a retrospective study with short-term follow-up, these results suggest that the posterior intercostal Volume < 73 30 29 19.5 7.7 0.0005 51.1 17.1 < lymph nodes may have prognostic significance. This data has served cm3 Volume > 0.0001 as a trigger for us to now routinely include the posterior intercostal 73 cm3 lymph nodes in our thoracic lymphadenectomies for malignant pleural No disease 22 37 24.8 10.3 0.0002 NR 19.2 < mesothelioma. Further investigation of this nodal station is indicated, and in fissures by 0.0001 it is likely that these nodes should be included in any future staging system CT Disease in for malignant pleural mesothelioma. fissures by CT Disclosure: No significant relationships. Normal 36 23 14.7 6.8 0.02 42.8 10.7 < Hemoglobin 0.0001 Anemia HIOC No HIOC 41 18 11.9 7.8 0.1 29.2 17.1 0.01 POSTER SESSION 1 September 12, 2012 11:30-12:30 Adjuvant CT 35 24 12.6 8.3 0.2 27.6 18.9 0.06 P1.21: PRE-OPERATIVE PROGNOSTIC ASSESSMENT No CT IDENTIFIES AN EXTREMELY FAVORABLE COHORT FOLLOWING # PLEURECTOMY WITH HYPERTHERMIC INTRAOPERATIVE Histology by biopsy On multivariate analysis, presence of disease in CHEMOTHERAPY (HIOC) FOR MALIGNANT PLEURAL the fissures was the only significant predictor for shorter TTR (HR 3.7, p=0.0004). Significant independent predictors of shorter OS included non- MESOTHELIOMA (MPM)

iMig2012.org • Abstract Book 90 epithelial histology (HR 2.9, p=0.02), volume > 73cm3(HR 2.2, p=0.04), disease in fissures by CT (HR 4.5, p=0.0006), anemia (HR 4.7, p<0.0001), and no treatment with HIOC (HR 2.6, p=0.008). For the 12 patients (20% of this cohort) with all five favorable predictors, the 5-year OS rate was 92%.

Conclusion: Presence of disease in the fissures on the pre-operative CT scan is a significant independent predictor for both early tumor recurrence and shorter survival among patients undergoing pleurectomy for MPM. Conversely, patients with epithelial histology on biopsy, normal hemoglobin levels, small volume disease and no fissure involvement have a remarkably long survival following pleurectomy and HIOC.

Disclosure: No significant relationships.

POSTER SESSION 1 September 12, 2012 11:30-12:30 poster sessions | september 12 P1.22: QUANTIFIED COMPUTED TOMOGRAPHY ASSESSMENT OF ASBESTOS-RELATED DISEASE

Carmen Endress1, Michael Harbut2, Anthony Endress1, Cynthia Noraian3 1Radiology, St John Providence Healthcare, MI/UNITED STATES OF AMERICA, 2Oncology, Karmanos Cancer Institute, Detroit/MI/UNITED STATES OF AMERICA, 3Oncology, Wayne State University, MI/UNITED STATES OF AMERICA

Background: The management of a patient with asbestos related disease starts with a correct diagnosis and determination of the extent of the disease. The radiographic diagnosis of asbestos related disease has largely depended on the B-Reader system, a diagnostic tool originally designed to study pneumoconiosis in large populations. This classification system is subjective with poor inter-observer agreement among experienced readers. Consequently, it has motivated us to seek a method of characterizing asbestos related disease utilizing current advances in technology and a novel reporting form to make a more confident diagnosis and depiction of asbestosis.

Methods: Our study of 50 patients was approved by the Institutional Review Board at Wayne State University and in accordance with HIPAA regulations. All participants provided signed informed consent. We outline our method to quantify and describe asbestos related disease based on computerized analysis of CT findings. An originally developed classification form based on computerized findings was used to record asbestos international mesothelioma interest group related abnormalities. The system provides a comprehensive evaluation of 3-dimensional (3D) rendering measurements of pleural plaques and surface area that will offer a reproducible way to measure and quantify asbestos related changes. Pulmonary fibrosis assessment is achieved using color segmentation; 3D analysis of nodules supplies size and shape determinations; lung segmentation volume is provided

Results: CT images with computer assisted software provided clarity, detection of subtle and advanced changes, and objective quantification of thoracic abnormalities in patients with asbestos related diseases. The newly created reporting form renders a device to comprehensively record and quantify the CT findings

Conclusion: We believe that our approach to classification and quantification of asbestos related abnormalities will help detect disease at an early stage and more clearly elucidate pathologies. Our expectation is to initiate a practical alternative to the B-Reader System for the classification of asbestos-related diseases and other types of pneumoconiosis.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 91 POSTER SESSION 1 September 12, 2012 11:30-12:30

P1.23: PATTERNS OF DISEASE PROGRESSION IN PATIENTS WITH MALIGNANT PLEURAL MESOTHELIOMA UNDERGOING PLEURECTOMY / DECORTICATION, HYPERTHERMIC PLEURAL LAVAGE WITH POVIDONE-IODINE FOLLOWED BY PROPHYLACTIC RADIOTHERAPY AND ADJUVANT CHEMOTHERAPY

Loic Lang-Lazdunski1, Andrea Bille’2, Lawrence Okiror2, Gary Cook3, James Spicer4, David Landau4 1Division Of Cancer Studies, King’s College London, United Kingdom/ UNITED KINGDOM, 2Department Of Thoracic Surgery, Guy’s Hospital, London/UNITED KINGDOM, 3Clinical Pet Centre, Division For Imaging Sciences, King’s College London/UNITED KINGDOM, 4Department Of Oncology And Hematology, Guy’s Hospital/UNITED KINGDOM

Background: There is no known cure for malignant pleural mesothelioma poster sessions | september 12 (MPM). Patients treated with multi-modality therapy generally experience longer survival. Time to progression and modes of relapse are well known following extrapleural pneumonectomy and adjuvant radiotherapy. We wished to evaluate the patterns of disease progression in patients treated with pleurectomy/decortication (P/D), hyperthermic pleural lavage with povidone-iodine, prophylactic radiotherapy and adjuvant chemotherapy.

Methods: Prospective study of patients treated with P/D and hyperthermic pleural lavage with povidone-iodine, prophylactic radiotherapy and adjuvant chemotherapy at our institution. Patients were followed up in our clinic regularly and had PET/CT within 6 weeks of completion of adjuvant chemotherapy and six-monthly thereafter. The first site of relapse on PET- CT was recorded and all scans were reviewed by an independent observer.

Results: Between October 2004 and March 2012, sixty five patients underwent P/D and hyperthermic pleural lavage with povidone-iodine, prophylactic radiotherapy and adjuvant chemotherapy. Thirty two patients (27 male, median age 61 year, range 45-73) had their PET-CT at our institution and were analyzed. Eighteen of 32 patients were alive at last follow-up (median follow-up 39.6 months, range 16-76 months). Nine patients were alive with no evidence of disease recurrence, 9 were alive with disease recurrence, of whom 6 patients experienced a local recurrence in the pleural cavity and one patient experienced a recurrence in the pleural cavity and in the mediastinum, 2 patients experienced a recurrence around their diaphragmatic mesh. Fourteen patients have died

of disease progression (median survival 24.7 months, range 15-38). Four international mesothelioma interest group patients experienced a local recurrence in the pleural cavity, two patients progressed in the pleural cavity and mediastinum, one patient had relapse in both pleural cavities, one patient progressed in a supraclavicular lymph node and two patients progressed in coeliac lymph nodes (N3), two patients progressed with bone metastases (vertebral metastases) and one patient presented with peritoneal metastases. Interestingly, no patient relapsed in the thoracotomy scar. . The median maximum standardized uptake value (SUVmax) in relapsing mesothelioma was 10.9 (range 4.9 – 27.3). There was a statistical correlation between the SUVmax of recurrent mesothelioma and survival (p=0.05).. In this group of patients, the median disease free survival (DFS) was 9 months (range 6-18). There was a statistical correlation between DFS and complete macroscopic resection (p=0.02) but there was no correlation between DFS and histologic subtype (epithelioid vs biphasic/sarcomatoid|).

Conclusion: After Pleurectomy/decortication with hyperthermic pleural lavage with povidone-iodine, prophylactic radiotherapy and adjuvant chemotherapy the most frequent site of recurrence is the pleural cavity. Peritoneal seeding is rare. Tumor SUVmax does not significantly correlate with survival.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 92 Poster Session 1 Pathology and Cytology September 12, 2012 11:30-12:30

POSTER SESSION 1 September 12, 2012 11:30-12:30 POSTER SESSION 1 September 12, 2012 11:30-12:30

P1.24: NATIONAL MESOTHELIOMA VIRTUAL BANK (NMVB): P1.25: HIGH PREVALENCE OF ATYPICAL MESOTHELIAL A PLATFORM FOR COLLABORATIVE RESEARCH AND PROLIFERATION IN EXTRAPLEURAL PNEUMONECTOMY MESOTHELIOMA TISSUE RESOURCE TO ENHANCE TRANSLATION SPECIMENS; FURTHER EVIDENCE OF A POTENTIAL PRECURSOR RESEARCH. LESION TO INVASIVE MESOTHELIOMA poster sessions | september 12

Waqas Amin1, Anil V. Parwani2, Nancy Whelan3, Rajiv Dhir2, Michael Leona Doyle1, Lucian Chirieac2 Feldman4, Jonathan Melamed5, Harvey Pass6, Raja Flores7, Michael J. 1Department Of Pathology, Brigham And Women’s Hospital, MA/UNITED Becich3 STATES OF AMERICA, 2Department Of Pathology, Brigham And Women’s 1Biomedical Informatics, University Of Pittsburgh, Pittsburgh/PA/ Hospital, Boston/MA/UNITED STATES OF AMERICA UNITED STATES OF AMERICA, 2Pathology, University Of Pittsburgh Medical Center, Pittsburgh/PA/UNITED STATES OF AMERICA, 3Biomedical Background: Atypical mesothelial proliferation (AMP) is thought to Informatics, University Of Pittsburgh, Pittsburgh/UNITED STATES OF represent a potential precursor lesion to invasive pleural mesothelioma. AMERICA, 4Pathology, University Of Pennsylvania, Philadelphia/PA/UNITED To our knowledge there is no published literature describing the STATES OF AMERICA, 5Pathology, New York University Medical Center, clinicopathologic characteristics of AMP. The aim of this study was New York/NY/UNITED STATES OF AMERICA, 6Cardiothoracic Surgery, to evaluate the prevalence of AMP in extrapleural pneumonectomy New York University Medical Center, New York/NY/UNITED STATES OF (EPP) specimens for invasive mesothelioma and to correlate AMP with AMERICA, 7Cardiothoracic Surgey, Mount Sinai School Of Medicine, New clinicopathologic features. York/UNITED STATES OF AMERICA Methods: We studied 46 consecutive EPPs with available surgical Background: The National Mesothelioma Virtual Bank (NMVB), developed material (mean 22 slides per case, range 12-30), performed for invasive six years ago, gathers clinically annotated human mesothelioma specimens mesothelioma (IM) over 16 months. Each case was assessed independently for basic and clinical science research. During this period of time, this by two pathologists for AMP according to currently established resource has greatly increased its collection of specimens by expanding morphologic criteria. We evaluated architectural and cytologic features, the number of contributing academic health centers. The resource the prevalence and extent of AMP and correlated clinicopathologic features has provided hundreds of high quality, well characterized annotated between mesotheliomas with and without AMP. biospecimen to the mesothelioma research community. Results: All 46 EPPs (40M/6F, mean age 62.9 years; range 38-79) Methods: The NMVB collaborating sites included New York University, showed invasive mesothelioma (n=30 epithelioid, n=15 mixed and n=1 University of Pennsylvania, and the University of Pittsburgh Medical sarcomatoid). AMP was identified in 10 (22%) EPP specimens, in a mean of Center; Mount Sinai School of Medicine joined the collaboration in 2011 3.5 slides (range 1-6). Nine cases (90%) were associated with epithelioid international mesothelioma interest group to increase the collection of specimens thereby allowing the resource to mesothelioma and 1 case (10%) with mixed mesothelioma. Common progress and meet the needs of research communities. Marketing efforts architectural patterns of AMP were a single cell layer proliferation (n=8), at both national and international annual conferences increase awareness stratified proliferations (n=5) and papillary proliferations (n=5). Six cases and availability of the mesothelioma specimens at no cost to approved (60%) had mixed AMP growth patterns. In AMP with a single cell layer investigators who query the web-based NMVB database for cumulative proliferation, prominent nucleoli were present in at least 50% of lesional and patient level clinicopathology information on the specimen. The NMVB cells. AMP was present in EPPs weighing less (median 747g vs. 1110g, resource includes three distinct Tissue Microarrays (TMAs) that encompass p=0.03) and in older patients (68 vs. 63 years, p=0.02). more than 100 malignant mesothelioma cases for biomarker research. Conclusion: In our study the prevalence of AMP (22%) in EPPs is higher Results: The NMVB database provides researchers real-time, interactive than anticipated, is more frequent in older patients and in specimens access to richly annotated specimens and essential information related to with lower weights. Further studies are needed to investigate the mesothelioma. The data disclosure and specimen distribution protocols clinical significance of AMP and the role of AMP in the pathogenesis of are tightly regulated to maintain compliance with participating institutions’ mesothelioma. IRB and regulatory committee reviews. The NMVB currently has over 1000 annotated cases available for researchers, including paraffin embedded Disclosure: No significant relationships. tissues, fresh frozen tissue, tissue microarrays, blood samples and genomic DNA. In addition, the resource offers expertise and assistance for collaborative research. POSTER SESSION 1 September 12, 2012 11:30-12:30 Conclusion: The National Mesothelioma Virtual Bank (NMVB) is a virtual biospecimen resource with robust translational biomedical informatics P1.26: MESOBANK - A UK BASED BIORESOURCE FOR MALIGNANT support to facilitate basic science, clinical, and translational research. MESOTHELIOMA Furthermore, in the last six years the resource has provided hundreds of fresh frozen, paraffin, blood samples and dozens of TMA slides to the Dean Fennell1, Stefan Marciniak2, John Edwards3, Peter Szlosarek4, research community. The investigators request specimens and/or data Doris Rassl2, Sam Janes5, Keith Kerr6, Peter Hamilton7, Victoria Hughes2, by submitting a Letter of Intent which is then evaluated by mesothelioma Robert C. Rintoul2, Richard Booton8 research evaluation panel. 1University Of Leicester, Leicester/UNITED KINGDOM, 2Papworth Hospital, Cambridge/UNITED KINGDOM, 3Dept Of Surgery, Northern General Disclosure: No significant relationships. Hospital/UNITED KINGDOM, 4Molecular Oncology, Barts Cancer Institute,

iMig2012.org • Abstract Book 93 Queen Mary University Of London, London/UNITED KINGDOM, 5Respiratory malignant mesothelioma of the pleura, there has been several guidelines Medicine, University College London/UNITED KINGDOM, 6Dept Of and recommendations given by various panel for making a definitive Pathology, University Of Aberdeen/UNITED KINGDOM, 7Health Sciences, diagnosis of malignant mesothelioma. The most recent is the “Guidelines University Of Belfast/UNITED KINGDOM, 8Respiratory Medicine, For Pathologic Diagnosis of Malignant Mesothelioma: A Consensus Wythenshawe Hospital/UNITED KINGDOM Statement From The International Mesothelioma Interest Group and International mesothelioma Panel”. They all recommand to perform an Background: It is now widely accepted that progress in the treatment of immnuhistochemical analysis and to use two positive and two negative mesothelioma is only likely to come with a better understanding of tumour markers before making a definitive diagnosis of mesothelioma.. But biology through integration of basic science and translational medicine there is no absolute antibodies that might definitively give a diagnosis approaches. Such work will be facilitated by the availability of quality- of mesothelioma. Moreover there are so many different antibodies assured, fully annotated tissue collected to rigorous standard operating from different sources, clones and techniques, that the pathologist procedures. Currently, few bioresources of mesothelioma tissue exist, may be confused and find difficult to find his way for a safe and secure the largest being the National Mesothelioma Virtual Bank hosted by the diagnosis of mesothelioma. We aimed to identify all literature related University of Pittsburgh (http://www.mesotissue.org/). Within the UK, to immunohistochemical markers tied to the diagnosis of malignant a few clinical/research groups hold fresh tissue from small numbers of mesothelioma. Search were conducted from the international literature mesothelioma patients but these collections are not formally linked and and relevant articles were identified and retrieved intended to improve the do not involve the collection of tissue and data in accordance with any diagnosis of malignant mesothelioma and its quality, to propose a strategy universal Standard Operating Procedures. and the best panel in term of sensibility and specificity and cut off of poster sessions | september 12 staining for selecting the most robuste markers on a routine practice level. Methods: The British Lung Foundation has recently confirmed funding for MesobanK, a UK based bioresource of malignant mesothelioma Methods: Meta-analysis was performed based on a literature review of tissue samples. This will comprise a) a retrospective collection of paraffin papers published in Pubmed and Medline electronic literature databases embedded tissue blocks to allow construction of a large tissue microarray between 1979 and 2010. The markers (EMA, HMFG-2, vimentin, keratin (1000 cases) and b) prospective collection of fresh frozen mesothelioma AE1/AE3, thrombomodulin, mesothelin, calretinin, HBME-1, WT-1, keratin tissue and matched blood samples (300 cases over 3 years) from multiple 5/6, n-cadherin, D2-40, and CEA, Ber-EP4, B72.3, CD15, MOC-31, centres across the UK. In addition, it is planned to develop 20 new fully e-cadherin, TTF-1, BG-8). Immunohistochemical markers were performed annotated mesothelioma primary cell cultures/cell lines. The bioresource on formalin-fixed paraffin-embedded tissues samples and markers will be supported by a web-based IT infrastructure for annotating and performed on cytology specimens were not analysed. Papers analyzing less searching the collection. Clinical data will be collected on each case and than 10 tumours, abstract or case report were excluded. The sensitivity, supplemented by laboratory and pathology results, Hospital Episode specificity and their 95% confidence intervals were calculated at 5 cut-offs Statistics data and Cancer Registry data in order to achieve as complete a of % of cells expression: 0%, 10%, 25%, 50% and 75%. data set as possible on each case. Results: According to the 99 selected papers, our results suggest that the Results: MesobanK will follow the Guiding Principles laid out by the NCRI most sensitive and specific panel to be used in the differential diagnosis Confederation of Cancer Biobanks and the Medical Research Council of EM from metastatic adenocarcinoma includes WT-1, keratin 5/6, Operational and Ethical Guidelines on Human Tissue and Biological monoclonal CEA, BG-8 and MOC-31 preferentially evaluated at 0% staining Materials for Use in Research. It will also be managed within the scope cells cut-off, thrombomodulin, D2-40 and B72.3 at 10% of tumour cells of all relevant regulatory frameworks and quality management/quality cut-off, calretinin and ber-EP4 at 25% cut-off. assurance systems. In addition, we share the aim of the US National Cancer Institute (NCI) National Biospecimen Network Blueprint: to create Conclusion: According to the recommendations of the International a comprehensive framework for sharing and comparing research results Mesothelioma Interest Group, the tumour cells staining can be credibly through a robust, flexible, scalable and secure bioinformatics system that evaluated from 1%, up to 10% cut-off for keratin 5/6, up to 25% cut-off for supports the collection, processing, storage, annotation and distribution of monoclonal CEA and up to 50% cut-off for calretinin, BG-8, ber-EP4 and biospecimens and data using standard operating procedures based on best MOC-31. international mesothelioma interest group practices. Disclosure: No significant relationships. Conclusion: A steering committee will be set up which will have overall control of MesobanK. An independent scientific advisory board will review applications for samples and advise the steering committee. Prioritisation for access to samples will be based solely on scientific merit. POSTER SESSION 1 September 12, 2012 11:30-12:30 All researchers, whether in the UK National Health Service, universities, charities, government agencies or commercial companies, and whether P1.28: MALIGNANT MESOTHELIOMA CAN IN MOST CASES BE based in the UK or abroad will be subject to the same application process CORRECTLY DIAGNOSED BASED ON EFFUSIONS. and approval criteria. It is anticipated that initial tissue samples will be available in late 2013 Anders Hjerpe1, Filip Mundt2, Katalin Dobra1 1Department Of Laboratory Medicine, Karolinska Institutet, Stockholm/ Disclosure: No significant relationships. SWEDEN, 2Laboratory Medicine, Karolinska Instiute, Stockholm/SWEDEN

Background: The IMIG recommendations state that the diagnosis of a malignant mesothelioma should be based on the examination of a biopsy, POSTER SESSION 1 September 12, 2012 11:30-12:30 obtained at thoracoscopy. Exfoliated MGG- or Pap-stained cells from an effusion is considered to be insufficient for the diagnosis. However, the P1.27: DIAGNOSIS OF PLEURAL EPITHELIOID MALIGNANT refinement of adjuvant techniques during the last decades has changed MESOTHELIOMA: A META-ANALYSIS BASED ON ROUTINELY this completely. USED IMMUNOHISTOCHEMICAL MARKERS. Methods: We intend to present our experiences with such analyses in Nolwenn Le Stang1, Marie Karanian2, Maria Paciencia2, Françoise effusion cytology. With the help of immunocytochemistry, FISH, biomarker Galateau-Sallé1 analyses and electron microscopy, we can establish the mesothelioma 1Pathology Department - University Hospital, Mesonat Registry (U 1086 diagnosis in a majority of cases. Inserm), Caen Cedex 9/FRANCE, 2Pathology Departement - University Hospital, U 1086 Inserm “Cancers & Préventions”, Caen Cedex 9/FRANCE Results: In a laboratory with experiences of these techniques, the diagnosis is accurate and definite, and will provide all information Background: Because of the well known heterogeneity and mimics of needed for choice of therapy in cases when surgery is not considered.

iMig2012.org • Abstract Book 94 The diagnosis can in this way be obtained already with the first effusion Prognosis:* withdrawn, and it will save these patients from a series of cumbersome examinations.

Conclusion: In these cases, when the diagnosis is definite, additional biopsy sampling must be considered unethical. We advocate that IMIG should change their recommendations, regarding how the diagnosis is made.

Disclosure: No significant relationships.

POSTER SESSION 1 September 12, 2012 11:30-12:30

P1.29: A REVIEW OF THE (SCARCE) LITERATURE ON LOCALIZED MALIGNANT MESOTHELIOMA poster sessions | september 12 Steven Kazan, Justin A. Bosl Kazan Mcclain Lyons Greenwood & Harley, Oakland/CA/UNITED STATES OF AMERICA

Background: Localized malignant mesotheliomas occur only rarely. The literature gives rise to two main questions: (1) What is the prognosis? Some authors suggest that surgical cure is possible, but many of the reported cases have gone on to die of the malignancy after the passage of some years. (2) Was there any history of asbestos exposure or of employment in trades where such exposure was likely? The paucity of cases makes * Charts prepared by Dr. Allan Smith (U.C. Berkeley School of Public Health) conducting a full epidemiological study complicated. to aid in illustrating his testimony in a legal case.

Methods: A literature search was conducted for cases of localized Conclusion: Among the localized malignant mesothelioma patients malignant mesothelioma to examine what asbestos exposure is reported for whom an exposure history was available, the rate of men that were and what the survival and followup have been. exposed falls within the range of exposure rates among diffuse malignant mesothelioma patients. The follow up in many cases with respect to Results: prognosis has been insufficient to justify any claims that these patients can be cured. Certainly, there is a significant number of patients that do die of Asbestos Exposure:* disease during the limited follow up reported.

Disclosure: No significant relationships. international mesothelioma interest group

iMig2012.org • Abstract Book 95 Poster Session 1 Cell Lines and Animal Models September 12, 2012 11:30-12:30

POSTER SESSION 1 September 12, 2012 11:30-12:30 POSTER SESSION 1 September 12, 2012 11:30-12:30

P1.30: DEVELOPMENT OF TRACT METASTASES AFTER P1.31: CHARACTERISATION OF A PRE-CLINICAL RAT MODEL OF CENTESIS PROCEDURE DIFFER BETWEEN MURINE MODELS OF MESOTHELIOMA MESOTHELIOMA

Amanda Hudson, Chris Weir, Rozelle Harvie, Elizabeth Moon, Stephen poster sessions | september 12 Robin Cornelissen1, J G J V Aerts2, M E.H. Lambers2, H C Hoogsteden2, Clarke, Nick Pavlakis Joost P.J.J. Hegmans2 Medical Oncology: Royal North Shore Hospital, Bill Walsh Cancer Research 1Pulmonary Medicine, Erasmus Mc, Rotterdam/NETHERLANDS, 2Pulmonary Laboratories, Sydney/NSW/AUSTRALIA Medicine, Erasmus Medical Centre, Rotterdam/NETHERLANDS Background: Malignant mesothelioma (MM) is a relatively rare and locally Background: Carcinogenesis along the tracts of cytology or biopsy aggressive tumour resulting in short survival and high morbidity. The needles, chest tubes, thoracoscopy trocars and surgical incisions is incidence of MM is increasing due to the widespread use of asbestos in the problematic complication in malignant mesothelioma. To prevent industrialised world, with Australia having the highest per capita incidence. malignant seeding at sites of diagnostic or therapeutic interventions, The majority of patients with MM are diagnosed in advanced disease prophylactic irradiation of tracts (PIT) was introduced in an attempt to stage when administration of systemic chemotherapy represents the main improve quality of life for these patients. However, the effect of PIT seems active treatment option hoping to extend survival and improve symptoms. dependent on the incidence of tract metastasis in the particular studies However due to the inherent resistance of this disease, relatively poor so no definitive conclusions can be drawn from the three randomized response to treatment is seen and relapse after initial chemotherapy is controlled trials that have been published so far. PIT will only be effective the norm. There is as yet no proven effective treatment after failure of in the subset of patients in which the tumor is prone to show growth after first line chemotherapy, and consequently there is an urgent need for a local intervention. We anticipate that the occurrence of tumor seeding the development of novel and more effective treatments which may at the puncture side is related to tumor characteristics. Genomic and also circumvent drug resistance. The development, therefore, of a well phenotypical features of individual differences between mesothelioma characterised clinically relevant animal model is paramount in order to test patients may influence their immunological and stromal components novel drugs and treatment strategies. Although the syngeneic Fischer 344 within the tumor environment. To investigate this concept, we studied the rat IL-45 mesothelioma cell line has previously been used as a model for effects of a needle puncture in the peritoneal cavity on tumor outgrowth of mesothelioma, little characterisation has been reported. Here we describe two well-defined mesothelioma cell lines in tumor bearing mice. The two the characterisation of the rat IL-45 mesothelioma cell line as a pre-clinical cell lines were selected for their known different growth pattern in mice. rat model of mesothelioma.

Methods: CBA/j mice and BALB/c mice (n=8) were inoculated Methods: IL-45 cells were grown using standard culturing techniques intaperitoneally with either an AC-29 or AB-1 tumor cell line, respectively. (37°C, 5% CO2 humidified incubator). Resistant cell lines were developed international mesothelioma interest group Tumor cells were injected at the left side of the abdomen. After 8 days a by repeatedly exposing parental IL-45 cells to known active agents peritoneal puncture was done at the contralateral side of the abdomen. (cisplatin, pemetrexed, gemcitabine and vinorelbine) and cell viability The mice were sacrificed on day 12 and the peritoneum was analyzed for assays (MTT) were used to determined resistance to drug therapy. RNA local tumor growth at the site of the puncture. was extracted and qRT-PCR performed to investigate gene expression levels. Western blot will confirm correlations between gene and protein Results: All mice inoculated with the AC-29 cell line showed in tract expression levels. In vivo experiments used 0.5 x 106 IL-45 cells injected into metastasis at the puncture side, while none of the AB-1 treated mice the flank or pleural cavity of Fischer 344 rats. IHC was used to determine developed tumor outgrowth after centesis Histological examination of the histological differences between parental and drug resistant IL-45 cells and tumor at the primary side showed marked tumor cells, stromal cells and tumours. an influx of inflammatory cell types. Gene expression profiling of the two different cell lines showed differences, which may explain this different Results: In vitro, IL-45 cells undergo a normal cell cycle in approximately growth pattern. 14 hrs and have a biphasic appearance. Drug resistant cells are ≥ 2-fold resistant to the drug they were treated with. qRT-PCR results looking at Conclusion: Differences in tumor cell genotype and phenotype, and the mechanisms used by these cells to overcome drug treatment suggest subsequent changes in recruited immunological cells, could be the key that ABC transporters, hormonal receptors, and retinoic acid binding to predict which patient will develop tract metastasis. Correlating these proteins are involved. In vivo, IL-45 cells grow as sarcomatoid tumours results to a clinical model is warranted to produce a tract metastasis with pleomorphic features. These tumours can be grown either in the prediction model. flank, giving a large window of opportunity to test novel treatments (approximately 40 days), or in the pleural cavity where they growth Disclosure: No significant relationships. extremely aggressively resulting in gross pleural extension, local invasion and rapid weight loss in approximately 15 days.

Conclusion: We believe that our pre-clinical model is now ready to be used as a platform for testing new drugs in order to treat this important disease.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 96 POSTER SESSION 1 September 12, 2012 11:30-12:30 SWITZERLAND, 2Division Of Oncolocy, University Hospital Zurich, Zurich/ SWITZERLAND, 3Laboratory Of Molecular Oncology, University Hospital P1.32: IPILIMUMAB ALONE OR IN COMBINATION WITH Zurich, Zurich/SWITZERLAND IMMUNOTHERAPY IN MURINE MALIGNANT MESOTHELIOMA Background: Multimodal treatment currently provides the best survival Lysanne A. Lievense, M E.H. Lambers, N M Mahaweni, H C Hoogsteden, outcome for malignant pleural mesothelioma (MPM); however, local tumor Joost P.J.J. Hegmans, J G J V Aerts recurrence remains a significant challenge. Activation of the hedgehog Pulmonary Medicine, Erasmus Medical Centre, Rotterdam/NETHERLANDS signalling pathway could be detected in MPM (see abstract Shi, Y., et al.). As stem cell signalling plays an important role in tumor recurrence and Background: Recently the immunostimulating agent ipilimumab was metastasis, we hypothesized that hedgehog activation is an important approved by US food and drug administration (FDA) based on the increase factor in MPM recurrence after surgical resection and chemotherapy. In this in overall survival of patients with metastatic melanoma, although adverse regard, a chemoresistant side population of cells which retain precursor events and side effect of treatment are substantial. Ipilimumab blocks properties has recently been identified in MPM (C. Frei, 2011). Hedgehog cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) which is present antagonists targeting and inhibiting the cell surface activator of hedgehog on activated T-cells. When CTLA-4 is expressed it limits their activation. signalling, smoothened (SMO) have already been utilised successfully in Therefore, blocking CTLA-4 with ipilimumab is assumed to sustain several clinical trials such as in combination with cisplatin/etoposide in antitumor T-cell responses. Based on this feature it may be regarded as an small cell lung cancer, recurrent or non-responding medulloblastoma. attractive immunostimulating agent to further increase the effectiveness of Moreover, a study in a colon cancer xenograft model showed that a poster sessions | september 12 immunotherapy approaches. Malignant mesothelioma is an immunogenic hedgehog antagonist could potentially suppress tumor recurrence (F. cancer with poor prognosis. We showed in previous preclinical and clinical Varnat, 2009). studies that dendritic cell-based immunotherapy is safe and feasible and induced cytotoxic T cell responses. Further boosting the antitumor immune Methods: In this study, we aim to use an in vivo rat bioluminescent MPM response with ipilimumab could be a valuable additive to our current model (Y. Shi, 2011) to study hedgehog antagonism in combination with immunotherapy strategy. The aim of this study is to investigate whether cisplatin as a potential means to prevent tumor recurrence. Cytotoxicity ipilimumab alone or in combination with immunotherapy is of importance and anchorage dependent colony formation assays, were applied to in a mesothelioma mouse model, justifying clinical introduction in determine sensitivity of rat mesothelioma cells to a hedgehog antagonist in mesothelioma patients. vitro. The expression analysis was performed by quantitative real time PCR and compared by ∆∆Ct method. To study the effect of hedgehog Methods: Immunocompetent BALB/c mice were inoculated with a lethal antagonist on bioluminescent imaging, rat bioluminescent MPM cells were dose of AB1 tumor cells intraperitoneally (i.p.) and monitored for a month. treated with hedgehog antagonist and the resulting photon signal was Without further treatment, mice develop first signs of terminal illness detected using a small animal imager, IVIS, immediately after applying between 2 and 4 weeks. First in a phase I and II protocol a safe and luciferase substrate. efficient dosage of ipilimumab (antibody 4F10 in mice) was determined in healthy and tumor bearing mice. In addition, this was also determined Results: Rat MPM cells express key molecules of hedgehog signalling during DC-immunotherapy. The optimal dosage was used in a study namely transmembrane hedgehog activator, smoothened (SMO) which comparing efficacy of ipilimumab in four groups; tumor bearing mice is a target of hedgehog antagonist and Gli1, a transcription factor which ongoing no treatment (n=6), DC-therapy (n=6), ipilimumab (n=6), or DC- is a positive modulator of the hedgehog signalling pathway. Hedgehog therapy plus ipilimumab (n=6). antagonist can suppress growth of rat mesothelioma cells as well as bioluminescent rat mesothelioma cells in a dose dependent manner. We Results: In the dose-finding experiments both 200 ug and 400 ug of 4F10, further defined that hedgehog antagonist did not interfere with our in lead to a survival benefit in tumor-bearing mice. However, 400 ug had vivo bioluminescent assay. toxicity in 25% of the healthy mice and therefore 200 ug was selected as the optimal dosage. The median survival after tumor injection was 15 Conclusion: These results implicate a role of the hedgehog signalling days in the control group, 15 days in the ipilimumab group, 21 days in the pathway in the rat recurrence mesothelioma model. Based on these international mesothelioma interest group DC therapy group and 19 days in the DC therapy group plus ipilimumab. findings in vivo studies will be carried out. The overall survival after 30 days was 0% in the control group, 17% in the ipilimumab group, 50% in the DC-therapy group and 0% in the DC- Disclosure: No significant relationships. therapy plus ipilimumab group. Kaplan-Meier analyses showed statistically significant differences in survival curves comparing DC-therapy with control (p<0.001, by log-rank test) and DC-therapy plus ipilimumab and control (p<0.05, by log-rank test). POSTER SESSION 1 September 12, 2012 11:30-12:30

Conclusion: In this study we demonstrate that ipilimumab alone has P1.34: INTRAPERITONEAL OR INTRATRACHEAL: WHICH ROUTE modest effects on overall survival in our murine mesothelioma model. As OF EXPOSURE IS RELIABLE FOR SCREENING MESOTHELIOMA shown from other studies, DC-based immunotherapy generates significant RISK OF FIBROUS DUSTS IN RATS? beneficial effects both in the median and overall survival. In contrast to what was expected theoretically, no additional beneficial effects were Miho Mizoi1, Shuichi Adachi2, Masahiro Maeda3, Okio Hino4 seen when murine ipilimumab was combined with DC treatment. We are 1Deptartment Of Public Health, Grad.Sch., Sagami Women’s University, currently testing the activity of different immunological cell types after Sagamihara, Kanagawa/JAPAN, 2Department Of Public Health, Sagami ipilimumab treatment. Women’s University, Sagamihara, Kanagawa/JAPAN, 3Immuno-Bilogical Laboratories., Co.,Ltd., Fujioka, Gunma/JAPAN, 4Department Of Molecular Disclosure: No significant relationships. Pathogenesis, Juntendo University, School Of Medicine, Tokyo/JAPAN

Background: A large number of fibrous materials including nanofibers have been developed and used not only as asbestos substitutes, but also POSTER SESSION 1 September 12, 2012 11:30-12:30 in various advanced technologies. Some of these fibers, particularly those with high durability and specific dimensions, have the potential to result in P1.33: PRELIMINARY CHARACTERIZATION OF HEDGEHOG the development of mesothelioma in experimental animals. A systematized INHIBITION IN A RAT MODEL OF MESOTHELIOMA RECURRENCE protocol to evaluate the carcinogenicity of new fibrous materials should be established to prevent mesothelioma cases caused by environmental Mayura Meerang1, Emanuela Felley-Bosco2, Byron Bitanihirwe1, Rolf exposures to hazardous fibrous dusts. We focused on identifying suitable Stahel3, Walter Weder1, Isabelle Opitz1 routes of exposure to rats in evaluation protocols. 1Division Of Thoracic Surgery, University Hospital Zurich, Zurich/

iMig2012.org • Abstract Book 97 Methods: We examined plasma N-fragment of expressed in renal findings support the potential utility of human mesothelioma cell lines as a carcinoma (N-ERC)/mesothelin levels in female F344 rats after tool for studying cellular, molecular and genetic aspects of the disease. intraperitoneal injection or intratracheal instillation of seven different types of fibrous materials. At 1, 3, 5 days and 1, 2, 4, 10 and 20 weeks after Supported by: The Prince Charles Hospital Foundation, Cancer Council administration, 30 μL of blood was collected from the tail vein. Plasma Queensland, Dust Diseases Board of NSW. N-ERC levels were measured using a Rat N-ERC/Mesothelin assay kit. Test materials were multi-wall carbon nanotube (MWCNT: mean length 3.0 Disclosure: No significant relationships. μm, mean width 0.10 μm), spherical TiO2 (P: mean diameter 0.27 μm), fibrous TiO2 (FT100: mean length 1.68 μm, mean width 0.13 μm; FT400: mean length >10 μm), silicon carbide whisker (SIC mean length 6.40 μm, mean width 0.30 μm), potassium titanate whisker (KT: mean length 6.0 POSTER SESSION 1 September 12, 2012 11:30-12:30 μm, mean width 0.35 μm), and chrysotile asbestos (Chr: mean width 0.109 μm). Mesothelioma induction has previously been observed after P1.36: XENOGRAFT DEVELOPMENT OF HUMAN MALIGNANT intraperitoneal injection of MWCNT, SiC, KT and Chr. PLEURAL MESOTHELIOMA IN THE IMMUNE DEFICIENT MICE AND ITS PROGNOSTIC VALUE ASSOCIATED WITH PATHOCLINICAL Results: Plasma N-ERC levels of rats administered MWCNT, SiC, KT or PARAMETERS OF PATIENTS chrysotile asbestos were continuously higher even at 10 weeks after i.p. than the other non-carcinogenic materials (P, FT100 and FT400). Licun Wu1, Ming Li2, Zhihong Yun1, Yidan Zhao1, Marc De Perrot1, poster sessions | september 12 Conversely, plasma N-ERC levels showed 1 peak after intratracheal Ming Tsao2 instillation, then diminished rapidly to control levels even with carcinogenic 1Latner Thoracic Surgery Research Laboratories, Toronto General Research fibers. Institute, University Of Toronto, Toronto/ON/CANADA, 2Ontario Cancer Institute, Princess Margaret Hospital, Toronto/ON/CANADA Conclusion: Since plasma N-ERC levels of rats with intraperitoneal injection of fibrous dusts showed a good correlation with carcinogenicity, Background: Xenograft models derived directly from malignant pleural particularly at 10 weeks after injection, this short-term animal experiment mesothelioma patients maintain essential features of the original tumors provides useful data for evaluating the mesothelioma risk of newly and make it possible to develop individualized therapy. The tumor bank of developed fibrous materials. xenografts could provide as a public resource of novel models to study this disease. We also report that engraftment in mice is a prognostic indicator Disclosure: No significant relationships. of outcome for the patients.

Methods: Fresh tumor tissues harvested from 50 MPM patients treated by extrapleural pneumonectomy (EPP), decortication or biopsy were POSTER SESSION 1 September 12, 2012 11:30-12:30 implanted subcutaneously into the immunodeficient mice. Xenografts were passaged up to five generations. At each passage tumor tissues were P1.35: ESTABLISHMENT AND CHARACTERIZATION OF SEVEN cryo-preserved for banking and fixed for histological examination. All HUMAN MALIGNANT MESOTHELIOMA CELL LINES patients were divided into 2 groups of xenograft development or failure in mice based on epithelioid histology, EPP or pre-operative chemotherapy. Vandana Relan1, Leanne Morrison1, Belinda Clarke2, Edwina Duhig2, Ian Response to cisplatin or/and pemetrexed was evaluated. A. Yang3, Kwun M. Fong3, Rayleen V. Bowman3 1Uq Thoracic Research Centre,, University Of Queensland, The Prince Results: 40% (20/50) of patients’ tumors developed xenografts and ten Charles Hospital, Brisbane/QLD/AUSTRALIA, 2Department Of Anatomical models have been passaged for five generations. The first generation Pathology, The Prince Charles Hospital/AUSTRALIA, 3Department Of models required 180±21 (70-344) days to reach the endpoints, while Thoracic Medicine, The Prince Charles Hospital, Brisbane/QLD/AUSTRALIA growth times from the second to fifth generation were 98±14, 94±11, 67±5, and 64±6 days, respectively. The overall survival of patients whose international mesothelioma interest group Background: Malignant mesothelioma is an aggressive tumour of serosal xenografts failed in mice is significantly better than those with xenograft surfaces most commonly pleura, usually caused by exposure to asbestos. development (p=0.02). Despite a lower rate of engraftment for epithelioid Characterised cell lines represent a valuable platform for discovery MPM (32.4%) compared to other types (61.5%), for 37 epithelioid patients, of molecular targets that could underpin new effective treatment for those whose xenografts failed in mice had significantly better survival than mesothelioma, improve diagnostics, and possibly determine pathways those whose xenograft developed (p=0.01). For 34 patients who received leading to effective secondary (post-exposure) prevention. EPP, patients with xenograft failure have slightly better survival, however, the difference is not significant (p=0.07). A similar result was obtained for Aim: To assess the biological characteristics of seven malignant 33 patients who had pre-operative chemotherapy (p=0.08). Preliminary mesothelioma cell lines derived from human pleural biopsy tissue or pleural evaluation of the models to cisplatin treatment demonstrated growth effusions. inhibition activity in 7/10 of xenografts, whereas pemetrexed did not result in significant benefit. Methods: Cells were established in tissue culture as adherent cell lines from small pieces of fresh resected pleural tumour tissue and pleural Conclusion: Primary MPM xenograft model might be a good model to effusion derived cell pellets. Mesothelial origin was assessed by standard test novel therapies, and an independent indicator for prognosis of MPM morphology, ultrastructure (Transmission Electron Microscopy) and patients. immunocytochemistry. Growth characteristics were assayed using growth curves and population doubling times. Cytogenetic analysis using spectral Disclosure: No significant relationships. karyotyping (SKY) was performed to assess chromosomal abnormalities. All cell lines were assessed for anchorage independent growth by soft agar colony assay.

Results: All seven-cell lines stained positive for calretinin and cytokeratin 19. Doubling time ranged from 30-72 hours. All cell lines exhibited numerical chromosomal abnormalities ranging from 41 to 113. Monosomy of chromosomes 8, 14, 22 or 17 was observed in five lines, and four different karyotypes were observed in one of the lines. All cell lines demonstrated capacity for anchorage independent growth.

Conclusion: Classic biological characteristics of mesothelioma are preserved in tumour derived cell lines maintained in vitro culture. These

iMig2012.org • Abstract Book 98 Poster Session 2 Surgery September 12, 2012 16:00-17:00

POSTER SESSION 2 September 12, 2012 16:00-17:00 POSTER SESSION 2 September 12, 2012 16:00-17:00

P2.01: THE IMPACT OF MACROSCOPIC COMPLETE RESECTION P2.02: COMPLICATIONS ASSOCIATED WITH RADICAL RADICAL PLEURECTOMY FOR MESOTHELIOMA ON PULMONARY PLEURECTOMY FOR MALIGNANT PLEURAL MESOTHELIOMA IN FUNCTION 77 PATIENTS poster sessions | september 12 Joseph Friedberg1, Melissa J. Culligan1, Mary Putt2, Benjamin French2, Joseph Friedberg1, Melissa J. Culligan1, Keith Cengel2, Charles B. Stephen M. Hahn3, Charles B. Simone3, James Stevenson4, Evan Alley5, Simone2, James Stevenson3, Evan Alley4, Daniel Sterman4, Eric J. Wang5, Keith Cengel3, Eric J. Wang6, Daniel Sterman5 Stephen M. Hahn2 1Thoracic Surgery, University Of Pennsylvania, Philadelphia/PA/ 1Thoracic Surgery, University Of Pennsylvania, Philadelphia/PA/UNITED UNITED STATES OF AMERICA, 2Biostatistics And Epidemiology, STATES OF AMERICA, 2Radiation Oncology, University Of Pennsylvania/ University Of Pennsylvania, Philadelphia/PA/UNITED STATES OF UNITED STATES OF AMERICA, 3Taussig Cancer Institute, Cleveland Clinic AMERICA, 3Radiation Oncology, University Of Pennsylvania/UNITED STATES Foundation, Cleveland/OH/UNITED STATES OF AMERICA,4Medicine, OF AMERICA, 4Taussig Cancer Institute, Cleveland Clinic Foundation, University Of Pennsylvania, Philadelphia/UNITED STATES OF Cleveland/OH/UNITED STATES OF AMERICA, 5Medicine, University Of AMERICA, 5Medical College Of Georgia/UNITED STATES OF AMERICA Pennsylvania, Philadelphia/UNITED STATES OF AMERICA, 6Medical College Of Georgia/UNITED STATES OF AMERICA Background: Initially employing extrapleural pneumonectomy as our preferred approach to achieve a macroscopic complete resection, our Background: Radical pleurectomy has become our approach for use of intraoperative photodynamic therapy (PDT) with its subsurface achieving a macroscopic complete resection for patients with malignant treatment effect motivated us to become increasingly aggressive with lung pleural mesothelioma. This procedure is being employed for all patients sparing surgery instead. Over the past several years, radical pleurectomy undergoing surgery-based treatment in our center, even in those has thereby emerged as our standard approach. This report details the with advanced stage cancer or bulky tumors. As an initial attempt to complications we have observed with this procedure. characterize the impact on quality of life this procedure has on the patients who receive it, we began to collect postoperative pulmonary function Methods: 103 patients underwent surgery for malignant pleural studies from them. This report compares these patients’ preoperative and mesothelioma, of which 77 underwent radical pleurectomy (including postoperative pulmonary function studies. the most recent 42 consecutive patients). This procedure has evolved, but it has always included the goal of achieving a macroscopic complete Methods: 13 patients (9 stage III, 4 stage IV) at a mean age of 61 (42-72) resection with preservation of the entire lung, phrenic nerve, and as underwent radical pleurectomy for malignant pleural mesothelioma. An much native diaphragm and pericardium as possible. All of the patients FEV1 value was recorded within one month preoperatively, and another underwent intraoperative adjuvant therapy after the radical pleurectomy was measured at a mean postoperative time of 15 (3-46) months. The (76 intraoperative PDT, 1 hyperthermic povidone iodine lavage). international mesothelioma interest group Wilcoxon test was then applied to these values. Results: Macroscopic complete resection was achieved in 76 out of 77 Results: The mean decrease in FEV1 was 0.18 (0.05-0.36) liters (p=0.066). patients (7/6/52/12 patients at Stages I/II/III/IV, respectively). Mean/ This corresponded to a mean decrease of 7.0% (4.5-9.0%) in the percent median length of stay was 13.5/14 days. Complications included: 3 predicted FEV1. thirty-day mortalities (1 readmission with tamponade/multiorgan failure, 1 subarachnoid hemorrhage, 1 aspiration pneumonia), 27 atrial fibrillations, Conclusion: This small retrospective series, acknowledging the limitations 3 pericardial effusions requiring drainage, 1 pulmonary embolism, 18 deep therein, revealed a postoperative decrease in FEV1 that did not achieve venous thromboses, 20 pneumonias (15 reintubations/8 tracheostomies), statistical significance. These radical pleurectomies were conducted in an 11 discharges with a Heimlich valve (0 reoperations), 5 chyle leaks, and 1 advanced stage cohort of 100% stage III or stage IV patients, a population diaphragm rupture. often considered candidates only for extrapleural pneumonectomy to achieve a macroscopic complete resection. The observed decrease in FEV1 Conclusion: Caution is indicated in interpreting this series by virtue of values following radical pleurectomies compares very favorably with what it reflecting: a single surgeon learning curve, the inclusion of patients has been reported following pneumonectomies. Because lung parenchyma from Phase I studies, the advanced nature of the cohort (83.1% stage III/ is preserved with a radical pleurectomy, we conjecture the decrease in IV) and, especially, the concomitant superimposed morbidity of PDT in FEV1 is likely related to a compromise in diaphragm mechanics. We feel 98.7% of the patients. Within that framework, the recorded mortalities this data is encouraging and will serve as a basis from which to study (3.9%), chyle leaks (6.5%), and single pulmonary embolism (1.3%) compare pulmonary function and quality of life assessments more comprehensively reasonably or favorably with other reports. Other complications occurred in patients undergoing radical pleurectomy for malignant pleural more frequently than what might be expected with surgery alone: atrial mesothelioma. fibrillation (35.0%), deep venous thrombosis (23.4%), pneumonia (26.0%), and reintubation (19.0%). It seems reasonable-to-likely that the addition Disclosure: No significant relationships. of PDT contributed to this complication rate. The single diaphragm rupture occurred after a primary repair of the residual native diaphragm muscle, and should likely have undergone patch repair, but could potentially have been adequate without the addition of PDT. At least 90% of the patients (all but several of the Stage I patients) had most or all of their visceral pleura removed in order to achieve a macroscopic complete resection, which would predictably result in large air leaks, with some being persistent. The 14.3% incidence of discharges with a Heimlich valve may also be related to the addition of PDT. Finally, the 3.9% incidence

iMig2012.org • Abstract Book 99 of tamponade (one patient who was already discharged, presented from upon these results we feel that it remains prudent to perform routine home a week later in extremis, and ultimately died within 30 days) was laparoscopy, to both rule out false positive and false negative studies, almost certainly related to PDT. These were recent complications for which even in the setting of both PET and CT scans. It also remains reasonable we have started to routinely fenestrate any preserved pericardial sacs. We to perform contralateral thoracoscopy on a selective basis, especially for conclude that radical pleurectomy can be done with acceptable mortality, patients who have not had a PET scan. but it has a higher incidence of certain transient morbidities, especially when accompanied by intraoperative PDT. Disclosure: No significant relationships.

Disclosure: No significant relationships.

POSTER SESSION 2 September 12, 2012 16:00-17:00

POSTER SESSION 2 September 12, 2012 16:00-17:00 P2.04: POST OPERATIVE ATRIAL FIBRILLATION AFTER EXTRA PLEURAL PNEUMONECTOMY: RISK FACTORS AND THE EFFECT OF P2.03: ROUTINE LAPAROSCOPIC AND SELECTIVE PROPHYLACTIC BETA BLOCKADE CONTRALATERAL THORACOSCOPIC STAGING IN 151 PATIENTS UNDERGOING SURGERY FOR MALIGNANT PLEURAL James Hardy1, Xiaoxia Liu1, David J. Sugarbaker2, Gyorgy Frendl3 MESOTHELIOMA 1Anesthesiology Perioperative And Pain Medicine, Brigham And Women’s poster sessions | september 12 Hospital, Boston/MA/UNITED STATES OF AMERICA, 2Division Of Thoracic Joseph Friedberg1, Melissa J. Culligan1, Gary Korus2, Jo Buyske2, Keith Surgery, Brigham And Women’s Hospital, Boston/MA/UNITED STATES OF Cengel3, Charles B. Simone3, James Stevenson4, Eric J. Wang5, Daniel AMERICA, 3Anesthesiology, Perioperative, And Pain Medicine, Brigham And Sterman6, Stephen M. Hahn3 Women’s Hospital, Boston/MA/UNITED STATES OF AMERICA 1Thoracic Surgery, University Of Pennsylvania, Philadelphia/PA/UNITED STATES OF AMERICA, 2Surgery, University Of Pennsylvania/UNITED Background: Atrial fibrillation (AF) is associated with increased mortality STATES OF AMERICA, 3Radiation Oncology, University Of Pennsylvania/ and morbidity following thoracic surgery. Patients undergoing extrapleural UNITED STATES OF AMERICA, 4Taussig Cancer Institute, Cleveland Clinic pneumonectomy (EPP) are at high risk for post operative AF. Prophylactic Foundation/UNITED STATES OF AMERICA, 5Medical College Of Georgia/ beta-blockers have been shown to reduce the incidence of post operative UNITED STATES OF AMERICA, 6Division Of Pulmonary, Allergy And AF after lung resection, but have not been studied following EPP. We aimed Immunology, University Of Pennsylvania School Of Medicine, Philadelphia/ to define the risk factors for AF following EPP. We tested our hypothesis UNITED STATES OF AMERICA that prophylactic beta-blockade reduces the incidence of AF after EPP.

Background: The primary oncologic consideration for offering surgery- Methods: 551 patients (>27y) underwent EPP for mesothelioma between based treatment for malignant pleural mesothelioma (MPM) is that 1998 and 2011 at our institution. Data for 354 patients were extracted the disease is confined to one hemithorax. Given the radicality and into our database through retrospective chart review, 29 patients nonstandard-of-care status of MPM surgery, our group has adopted a very were excluded for preexisting AF. Patients were monitored with cardiac aggressive approach to invasive staging. We routinely perform laparoscopy telemetry, and the presence of post operative AF (irregularly irregular and selective contralateral thoracoscopy to supplement radiographic heart rate lasting >15min) was recorded. Two patient cohorts: those who staging. This report is a result of those results. received prophylactic beta-blockade (245/354) and those who did not (108/354) were compared in our analysis. Prophylactic beta-blockade was Methods: 151 patients with MPM were considered candidates for different introduced into our protocol in 2002. Univariate and multivariate analyses surgery-based multimodal treatments. All patients had chest CT scans, were conducted to identify risk factors for the development of AF. 17 had MRI scans and 95 had PET scans. 13 patients were excluded from having laparoscopy due to histories of fused abdomens from prior Results: The incidence of AF was 49.7% (178/354). The time course of conditions. 139 patients underwent laparoscopy (typically two 5mm ports) post op AF is shown in Figure 1. 66 (61.1%) patients without prophylactic international mesothelioma interest group with peritoneal lavage and selective biopsies. 16 patients underwent beta-blockade developed post operative AF vs. 109 (44.5%) of those contralateral thoracoscopy (a single 1cm incision), based upon suspicion by on prophylactic beta-blockade (p=0.0040). Univariate analysis showed the radiologist and/or multidisciplinary team. 15/16 had thoracoscopy and that older age (p<0.001), right sided surgery (p=0.0028), and lowest laparoscopy and one had thoracoscopy alone. magnesium level on post op days 0-3 (p=0.013) were risk factors. Multivariate analysis showed that prophylactic beta-blockers can Results: There were no operative complications. 119 laparoscopies were significantly reduce the risk of AF (RR: 0.582; 95% CI: 0.452-0.749). In outpatient procedures, 5 were admitted overnight (urinary retention), 15 contrast, age (RR: 1.04; 95% CI: 1.026-1.056), preoperative (home) beta- who also had thoracoscopy were admitted overnight, 1 thoracoscopy alone blocker use (RR: 1.519; 95% CI 1.151-2.004), right sided surgery (RR: 1.290; was admitted overnight. Laparoscopy revealed 6 false positive studies (1 95% CI 1.105-1.639) increase the risk of AF. interpretation of diaphragm transgression and 5 metastatic deposits) and 7 false negative studies (radiographically occult metastases - 3 by lavage Conclusion: Patients undergoing EPP have a high rate of postoperative AF. and 4 by biopsy). All of the false positive and all of the false negative Reasons specific to this patient population may include surgical stripping of studies occurred in patients who had PET scans. 2 patients had PET and the pericardium, right heart strain following the division of the right or left CT positive findings that were confirmed by laparoscopy. 4/16 patients pulmonary artery and large fluid shifts. The introduction of prophylactic interpreted as having contralateral thoracic disease on CT alone were all beta-blockers was associated with a statistically and clinically significant false positives, 1/16 with a positive PET-CT was a true positive and 1/11 reduction in AF. Right sided EPP involves routine patch reconstruction of with a negative PET-CT, but thought suspicious by the multidisciplinary the pericardium which may explain the higher risk for AF in this subgroup. team, was a false negative. The increased risk of AF in patients taking beta blockers at the time of admission may be related to withdrawal of beta-blockers. References Conclusion: Radiographic staging of the abdomen was inaccurate in Jakobsen et al, J Cardiothorac Vasc Anesth 1997;11:6, 746-51 9.3% of the patient, 5.0% false negative and 4.3% false positive, using the 139 patients undergoing laparoscopy as the denominator. All of the Disclosure: No significant relationships. inaccuracies occurred in patients with both PET and CT. Contralateral chest staging was primarily inaccurate in the setting of false positive interpretations, with 4/5 (80%) proving biopsy negative. All four false positives were CT alone and the one true positive was also PET positive, but there was also one false negative PET-CT. Overall, using the 16 patients who underwent contralateral thoracoscopy as the denominator, radiographic staging was inaccurate in 5/16 (31%) of the patients. Based

iMig2012.org • Abstract Book 100 POSTER SESSION 2 September 12, 2012 16:00-17:00 prognosis and there is no consensus on standard therapy. Best survival data with a median survival had been reported after multimodality P2.05: CLINICAL IMPACT OF POST OPERATIVE ATRIAL treatment including extrapleural pneumonectomy (EPP), but MARS trial was FIBRILLATION FOLLOWING EXTRA PLEURAL PNEUMONECTOMY a negative report. We considered whether EPP was effective therapy for MPM. James Hardy1, Xiaoxia Liu1, David J. Sugarbaker2, Gyorgy Frendl3 1Anesthesiology Perioperative And Pain Medicine, Brigham And Women’s Methods: Between 2000 and 2011, 24 MPM patients were definitive Hospital, Boston/MA/UNITED STATES OF AMERICA, 2Division Of Thoracic diagnosed by thoracoscopic biopsy in our hospital. We analyse 19 patients Surgery, Brigham And Women’s Hospital, Boston/MA/UNITED STATES OF of epithelial and biphasic type MPM, exclude 5 of 24 were sarcomatoid AMERICA, 3Anesthesiology, Perioperative, And Pain Medicine, Brigham And type MPM. Women’s Hospital, Boston/MA/UNITED STATES OF AMERICA Results: Of the 18 men and one woman, the median age was 69.7 years Background: Atrial fibrillation (AF) is common following thoracic surgery, (range; 44-83). Thirteen patients were epithelial type and 6 were biphasic and is associated with increased mortality, morbidity and hospital length type. 6 patients (4 epithelial type and 2 biphasic type) underwent EPP. of stay. Extrapleural pneumonectomy (EPP) carries a high risk of morbidity, 10 (7 epithelial type and 3 biphasic type) of 13 no-EPP patients were including post operative AF. Surgery involves violation of the pericardium, performed chemotherapy. 3 (2 epithelial type and one biphasic type) significant hemodynamic changes and fluid shifts. The aim of this study of 13 were no treatment. One of 6 EPP patients was right side MPM and was to describe the association between the development of post 12 of 13 no-EPP patients were right side MPM. 5 of 6 EPP patients were poster sessions | september 12 operative AF and outcomes in this patient group. cStageII and one was cStageIII. 3 of 13 no-EPP patients were cStageIb, 4 were cStageII, 5 were cStageIII and one was cStageIV according to IMIG. Methods: 551 patients (>27yrs) underwent EPP for mesothelioma One of 3 no treatment patients was cStageIb, one was cStageIII and one between October 1998 and May 2011 at our institution. Data for 354 was cStageIV. 7 of no-EPP patients had platinum-based chemotherapy. patients were extracted into our database through retrospective chart 4 of 6 EPP patients were performed preoperative chemotherapy and two review. 29 patients were excluded for having a history of AF. Patients were were performed postoperative chemotherapy. 4 of 6 EPP patients were monitored postoperatively in the intensive care unit or step down unit up staging (II→III) after surgery, but there was no lymphnode metastasis. with continuous cardiac telemetry, and the presence or absence of post None of EPP patients received hemithoracic radiotherapy after surgery. operative AF was recorded. AF was defined as an episode of irregularly One of 6 EPP patients had empyema without fistula and another one had irregular heart rhythm lasting at least 15 min. Hospital length of stay, atrial fibrillation as postoperative complications. EPP related morbidity and ICU length of stay, time to extubation, requirement for reintubation, mortality were 33% and 0%. The overall 1-, 2- and 3-year survival after occurrence of post operative stroke, acute kidney injury, requirement for diagnosis for all EPP cases and no-EPP cases who performed chemotherapy renal replacement therapy, in-hospital mortality and 30 day mortality were were 100%, 80%, 40% and 80%, 46.7%, 11.7%. Median survival was 35.7 recorded. Univariate and multivariate Logistic regression analyses were months for the EPP group and 23.4 months for the no-EPP group. used to explore the association between AF and the outcomes. Conclusion: The EPP group in our hospital was the better prognosis than Results: The cohort of patients who developed postoperative atrial no-EPP group. We judged that there was no indication of EPP in a patient fibrillation was compared to those without AF. The incidence of AF peaked of right side MPM more than 70 years old and a patient of MPM 75 years or on post operative days 2 and 3. Post operative atrial fibrillation was older. It was thought that if we will be able to choose the patient of MPM, significantly associated with increased length of hospital stay 20 days EPP will be effective therapy for epithelial and biphasic type MPM. vs. 11.7 days(p<0.0001), increased length of ICU stay 11.9 days vs. 4.4 days (p<0.0001), increased in-hospital mortality 7.3% vs. 1.1%(p=0.006) Disclosure: No significant relationships. and increased likelihood of reintubation 30.4% vs. 6.0% (p<0.0001). The presence of post operative atrial fibrillation was not statistically significantly associated with increased time to initial extubation, 30 day mortality, incidence of acute kidney injury, requirement for renal POSTER SESSION 2 September 12, 2012 16:00-17:00 international mesothelioma interest group replacement therapy, or increased incidence of postoperative stroke. Multivariate analysis demonstrated that AF was a risk factor for in P2.07: A CLINICAL STUDY OF 5 EPITHELIAL TYPE DIFFUSE hospital mortality (p=0.024), independent of age, gender, left ventricular MALIGNAT PLEURAL MESOTHELIOMA PATIENS WHO function, history of diabetes, hypertension, use of intraoperative heated UEDERWENT EXTRAPLEURAL PNEUMONECTOMY chemotherapy, or use of intraoperative blood products. Hideyuki Kobayashi1, Osamu Kawamata2 Conclusion: EPP is one of two surgical interventions available for 1Surgical Department, Onomichi Municipal Hostpital, mesothelioma, at the expense of significant morbidities. Patients are at Onomichi hiroshima/JAPAN, 2Surgery, Onomichi Municipal Hospital, high risk for the development of post operative AF. AF was associated Onomichi/JAPAN with a statistically and clinically significant increase in ICU and hospital length of stay, and in-hospital mortality. Although the development of Background: Diffuse malignat pleural mesothelioma (MPM) has poor post operative AF may be a marker for other underlying physiological and prognosis and there is no consensus on standard therapy. The best survival inflammatory changes following EPP, it is a key modifiable risk factor. data with a median survival had been reported after trimodality therapy The use of the most effective prophylactic pharmacotherapy should be including Extrapleural pneumonectmy (EPP),but Mars trial was a negative considered in these patients. report. We considered whether EPP was effective therapy for MPM.

Disclosure: No significant relationships. Methods: Between 2006 and 2010, 15 patients were definitive diagnosed by thoracoscopic biopsy as epithelial type MPM, and 5 of 15 underwent EPP in our hospital. We analysed these EPP cases.

POSTER SESSION 2 September 12, 2012 16:00-17:00 Results: ALL cases were males, the median age 68 year (range;60-73).2 cases had right side MPM,3 cases had left side MPM. 2 cases were P2.06: A CLINICAL STUDY OF 19 EPITHELIAL AND BIPHASIC TYPE cStageⅡ,3 cases were cStage Ⅲ according to IMIG. 3 patients were DIFFUSE MALIGNANT PLEURAL MESOTHELIOMA PATIENTS received preoperative chemotherapy, 1 patient received postoperative chemotherapy, 1 patient was pre-postoperative chemotherapy; 3 Osamu Kawamata cases were CDDP + Pemetrexed,2 cases were CDDP + Gemcitabin. Surgery, Onomichi Municipal Hospital, Onomichi/JAPAN All cases underwent resection diaphragm and 3 cases underwent resection pericardium. 1 of 5 cases had atrial fibrillation as postoperative Background: Diffuse malignant pleural mesothelioma (MPM) has a poor complication. None of all received hemithoracic radiotherapy after surgery.

iMig2012.org • Abstract Book 101 The survival term after EPP were 21.1 months(death of cancer), 33.3 in other two patients with duration of 11 and 19 months after onset, months (death of cancer), 39.6 months (death of cancer), 20.3months respectively. bronchoscopic treatment*: (1) spraying basic fibroblast growth (death of cancer), 31.8 months (on survival).2 of 5 cases made reccurence factor, (2) submucosal injection of OK432, (3) spraynig fibrin glue on the opposite lung and pleura, and respiratory failure progressed rapidly by pleural effusion but no EPP cases didn’t made opposite reccurence. Conclusion: Although symptomatic BPF after EPP requires immediate surgical intervention, asymptomatic BPF may be conservatively observed. Conclusion: EPP cases had no major critical postoperative complication and were 100% on 1 year survival, therefore it indicated that EPP was Disclosure: No significant relationships. effective therapy. We also consider it is necessary for maintaining QOL to prevent MPM from spreading across to the other side on EPP case.

Disclosure: No significant relationships.

POSTER SESSION 2 September 12, 2012 16:00-17:00

P2.08: SILENT BRONCHOPLEURAL FISTULA FOLLOWING poster sessions | september 12 EXTRAPLEURAL PNEUMONECTOMY FOR MALIGNANT PLEURAL MESOTHELIOMA

Nobuyuki Kondo1, Fumihiro Tanaka2, Yoshitomo Okumura3, Seiji Matsumoto1, Teruhisa Takuwa1, Masaki Hashimoto1, Hayato Orui1, Ayumi Kuroda1, Shunichi Fukuda3, Noriaki Tsubota4, Kazuya Fukuoka5, Takashi Nakano6, Seiki Hasegawa1 1Thoracic Surgery, Hyogo College Of Medicine, Nishinomiya/ JAPAN, 2Second Department Of Surgery, University Of Occupational And Environmental Health/JAPAN, 3Thoracic Surgery, Itami City Hospital, Itami/ JAPAN, 4Thoracic Oncology, Hyogo College Of Medicine, Nishinomiya/ JAPAN, 5Cancer Center, Hyogo College Of Medicine, Nishinomiya/ JAPAN, 6Division Of Respiratory Medicine, Department Of Internal Medicine, Hyogo College Of Medicine, Nishinomiya/JAPAN

Background: Bronchopleural fistula (BPF) after pneumonectomy is usually accompanied with typical and very severe clinical symptoms, and rapidly leads to fatal condition. However, in some rare instances, BPF continues to be totally asymptomatic with only radiological sign of decreased fluid level in the ipsilateral chest.

Methods: BPF was diagnosed in 5 out of 45 patients in whom EPP was completed in Hyogo College of Medicine between July 2004 and March 2012. Two patients with distinct clinical symptoms were surgically repaired immediately after onset. In the remaining 3 asymptomatic cases, diagnosis of BPF was made upon bronchoscopy and bronchography (n=1) or upon international mesothelioma interest group partial emptying of postpneumonectomy space on chest X-ray (n=2). The interval between EPP and onset of BPF ranged from 18 days to 2 years.

patient age Induction EPP Bronchial postopretative Onset of bronchoscopic bronchograpy Treatment Outcome Therapy Stump RT emptying findings (after onset) Coverage (after EPP) 1 49 PEM/ left Yes no 18 days pin-hole at minor leakage bronchoscopic intrathoracic air CDDP bronchial stump treatment* (1) disappeared x3, (2)x2, (3)x3 (6 months) 2 56 PEM/ left Yes hemithoracic, 25 months none no leakage bronchoscopic persistent CDDP 54Gy treatment (1)x2 (19 months) 3 48 PEM/ right Yes hemithoracic, 111 days small air bubble no leakage bronchoscopic persistent CDDP 54Gy to and fro, staple treatment (1)x 1 (11 months) exposed

Results: Three patients with silent BPF were remained to be afebrile, asymptomatic, with no abnormal findings of blood tests throughout their clinical courses. Emptying of postpneumonectomy space on chest X-ray was seen 18 days, 111 days, and 2 years after EPP, respectively. Bronchoscopy and bronchography were performed immediately after emptying, and revealed very small BPF (n=1), to and fro movement of air bubble with exposed staples (n=1), or only unremarkable findings (n=1). All three patients were conservatively observed with only intrabronchial spraying of basic fibroblast growth factor with or without submucosal injection of OK-432 and spraying of fibrin glue. Emptying was disappeared 5 months after onset in one patient with bronchgraphy- proven BPF. Persistent emptying with no clinical symptoms was observed

iMig2012.org • Abstract Book 102 POSTER SESSION 2 September 12, 2012 16:00-17:00

P2.09: OPERATIVE INDICATION OF EXTRAPLEURAL PNEUMONECTOMY FOR MALIGNANT PLEURAL MESOTHELIOMA

Satoru Moriyama1, Motoki Yano1, Hidefumi Sasaki1, Yu Hikosaka1, Katsuhiro Okuda1, Masayuki Shitara1, Masayuki Tanahashi2, Haruhiro Yukiue2, Hiroshi Haneda2, Eriko Suzuki2, Naoko Yoshii2, Hiroshi Niwa2, Yoshitaka Fujii1 1Oncology, Immunology, And Surgery, Nagoya City University Graduate School Of Medecal Sciences, Nagoya/JAPAN, 2Thoracic Surgery, Respiratory Disease Center, Seirei Mikatahara General Hospital, Hamamatsu/JAPAN

Background: The role of extrapleural pneumonectomy (EPP) for patients with malignant pleural mesothelioma (MPM) has been controversial.

Methods: We retrospectively reviewed the records of 56 patients with MPM to determine the efficacy of operative procedures. These patients poster sessions | september 12 were considered as having resectable disease using preoperative radiological and physiological examinations.

Results: There were 44 males and 12 females, with a mean age of 64 years old. A past history of exposure to asbestos was identified in 20 patients (35.7%). Thoracoscopic pleural biopsy was the most effective method for diagnosis in 45 patients. It required 3.3 months (median) from the first visit to establish the diagnosis. In histological types, 35 (62.5%) were epithelial, 11 (19.6%) were biphasic, and 10 (17.9%) were sarcomatous. According to the IMIG clinical staging, 11 patients (19.6%) were diagnosed as stage I, 23 (41.0%) stage II, 18 (32.1%) stage III, and 4 (7.1%) stage IV. Thirty-five (62.5%) of 56 patients underwent surgical procedures, including EPP in 33 patients (58.9%) and pleurectomy/ decortication (P/D) in 2 (3.6%). Microscopically complete resection by EPP was achieved in 22 patients (39.3%). Preoperative chemotherapy was given in 3 patients. Intraopeartive hyperthermo-chemotherapy was added in 7 patients. Postoperative chemotherapy was given in 4 patients, radiotherapy in 10, both chemotherapy and radiotherapy in 2, and hyperthermo-chemotherapy in 6. Major postoperative complications occurred in 12 patients (36.4%) and thirty-day mortality was 9.1% (3/33) in patients with EPP. No patient who underwent P/D had operative complications or mortality. Nonsurgical therapy was selected in 21 patients (37.5%); chemotherapy in 13 patients (23.2%), radiotherapy in 3 (5.4%), hyperthermo-chemotherapy in 1 (1.8%), and palliative care alone in 4 (7.1%). The median and 5-year overall survival rates among all patients were 21 months and 19%, respectively. There was no significant difference international mesothelioma interest group in survival between the patients with or without operation. The median and 5-year survival rates who underwent surgery were 21 months and 24%, respectively, whereas those with nonsurgical therapy were 20 months and 0%, respectively. A univariate analysis demonstrated that complete resection by EPP (p = 0.022), epithelial histology (p = 0.001), and c-stage I-II disease (p = 0.003) were significant positive prognostic factors. The patients who achieved complete resection by EPP were more likely to have c-stage I-II (p = 0.002) and epithelial diseases (p = 0.024).

Conclusion: EPP should be selected for the patients with c-stage I-II and epithelial disease.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 103 Poster Session 2 Novel Therapeutics September 12, 2012 16:00-17:00

POSTER SESSION 2 September 12, 2012 16:00-17:00 and tumor samples. Using several mesothelioma cell lines, our group has previously shown that the small molecule inhibitor SU11274 specifically P2.10: COMBINED ANTISURVIVIN-CISPLATIN TREATMENT OF targets MET with an IC50 value of 2 to 3 µM. We have also reported several MALIGNANT PLEURAL MESOTHELIOMA unique mutations of MET in mesothelioma tumors. In the current study, we evaluated tivantinib (Arq197, ArQule), an oral, synthetic, non-ATP-

Julija Hmeljak, Maja Cemazar, Andrej Coer dependent competitive, small molecule inhibitor of MET in mesothelioma poster sessions | september 12 Faculty Of Health Sciences, University Of Primorska, Izola/SLOVENIA tumors and cell lines.

Background: Survivin (BIRC5) is a cancer specific apoptosis inhibitor and Methods: Genomic DNA was extracted from 13 fresh frozen mesothelioma a promising target for novel anticancer therapies, especially for tumors tumor tissue samples and MET was sequenced using exon specific primers. which presently show poor response to conventional treatment, such as The non-malignant mesothelial cell line Met5A, and the mesothelioma malignant pleural mesothelioma (MPM). MPM rarely responds to cisplatin- cell lines H2596, H513, H2052, H2461 and H28 were purchased from or radiation-based treatment and median survival remains around 1 year the American Type Culture Collection (Manassas,VA). Cell viability was after diagnosis, despite recent advances in treatment. This fact, added to evaluated using the standard Alamar blue assay. Briefly, exponentially increasing incidence, underlines the need for novel treatment options for growing cells were seeded in 96-well tissue culture plates in 100ul (10% MPM. Our previous retrospective study found that all MPM tissue samples RPMI). After 24 hr of incubation the cells were treated with tivantinib at the analysed expressed high levels of survivin, so the aim of the present study indicated concentrations in 1% RPMI. After 72 hours, Almar blue was added was to explore the therapeutic potential of survivin inhibition combined and fluorescence was read at 530/590 nM. Data was normalized to the with hypotonic cisplatin perfusion of survivin- expressing MPM cells in vitro. cells without treatment. IC50 values were calculated using Prism software.

Methods: Two MPM cell lines (H2052 and 211H) and an immortalized Results: We observed a dose-dependent decrease of cell viability in the mesothelial line (MeT-5A) were used for in vitro experiments. Survivin mesothelioma cell lines treated with tivantinib; the control Met5A cells expression was silenced by Stealth® siRNA lipofection. Lipofected cells were not sensitive at the dose range tested. A dramatic reduction in were treated with hypotonic cisplatin solutions (0, 1 or 10 μg/mL) for 15 cell viability in response to tivantinib was noted in the H2596 and H513 minutes 24 hours after lipofection. Effects of each single therapeutic mesothelioma cell lines that harbor the MET T1010I mutation compared approach and their combination on cell survival and proliferation were with H28 cells, which express wild-type MET, and the control Met5A cells. assessed with the clonogenic and MTS assays. Mutational analysis identified the previously reported juxtamembrane domain mutation (R970C) in 1 of the 13 mesothelioma tumor samples. Results: We found that either hypotonic cisplatin perfusion or survivin silencing significantly reduced survival and proliferation of all three Conclusion: Exposure to tivitanib resulted in the inhibition of proliferation cell lines tested when applied as single-agent approaches (p<0.05). of MET–expressing mesothelioma cell lines. This inhibition was most

But when both treatments were combined, strongly synergistic effects prominent in cell lines which harbored mutations in MET. We believe that international mesothelioma interest group were observed, since more than 90 % reduction in both survival and MET may be an appropriate therapeutic target in mesothelioma. An NCI- proliferation was achieved after application of only 1 μg/mL cisplatin to all sponsored phase II trial of tivantinib in previously treated mesothelioma three lipofected cell lines (p<0.05). patients is in development at the University of Chicago.

Conclusion: Our in vitro results confirm that a combined antisurvivin Disclosure: No significant relationships. and classic chemotherapeutic approach might be a viable direction for development of effective therapies for MPM. Both anticancer gene therapy and hypotonic intrapleural chemotherapy are feasible treatment options for MPM patients and their combination should be further explored. POSTER SESSION 2 September 12, 2012 16:00-17:00

Disclosure: No significant relationships. P2.12: TARGETING ESTROGEN RECEPTOR β FOR TREATMENT OF PLEURAL MALIGNANT MESOTHELIOMA

Giulia Pinton1, Arcangela Gabriella Manente1, Antonio Daga2, Luciano POSTER SESSION 2 September 12, 2012 16:00-17:00 Mutti3, Laura Moro4, Stefan Nilsson5 1Dept. Of Pharmacological Sciences, University Of Piemonte Orientale A. P2.11: INHIBITION OF THE MET RECEPTOR TYROSINE KINASE IN Avogadro, Novara/ITALY, 2Irccs San Martino-Ist, Genova/ITALY, 3Dept. MALIGNANT MESOTHELIOMA Of Medicine, Vercelli Hospital, Vercelli/ITALY, 4Dept. Of Pharmaceutical Sciences, University Of Piemonte Orientale “A. Avogadro”, Novara/ Rajani P. Kanteti, Immanuel Dhanasingh, H.L. Kindler, Ravi Salgia ITALY, 5Karo Bio Ab, Huddinge/SWEDEN Department Of Medicine, The University Of Chicago, Chicago/UNITED STATES OF AMERICA Background: The role of estrogen receptor (ER) β as mediator of anti- proliferative responses has been well documented in numerous scientific Background: The MET receptor tyrosine kinase (RTK) plays an important articles and selective activation of ERβ with an agonist has shown in role in a variety of malignancies, especially malignant mesothelioma. The vivo anti-tumorigenic efficacy in various animal tumor models. MET proto-oncogene encodes a trans-membrane tyrosine kinase receptor for HGF, a multifunctional protein involved in tissue repair and metastasis. Methods: We have explored the anti-proliferative activity of the highly MET contains a semaphorin domain at the N-terminus, a juxtamembrane selective ERβ agonist, KB9520, in human mesothelioma cell lines in domain, and a tyrosine kinase domain at the C-terminus of the protein. vitro and in mesothelioma mice models in vivo and studied its anti- MET and its ligand HGF are over-expressed in mesothelioma cell lines tumorigenic efficacy.

iMig2012.org • Abstract Book 104 Results: We have demonstrated that KB9520 treatment inhibits POSTER SESSION 2 September 12, 2012 16:00-17:00 propagation of the human ERβ positive REN mesothelioma cell line in culture by blockade of the cell cycle at G1. Introduction of ERβ into P2.14: IN VITRO SCREENING OF AN APPROVED DRUG LIBRARY the non-ERα and ERβ expressing human mesothelioma cell line MSTO- IDENTIFIES AGENTS WITH REPURPOSING POTENTIAL FOR 211H, sensitizes these cells for the anti-proliferative effect of KB9520. MALIGNANT MESOTHELIOMA Selective activation of ERβ with KB9520 inhibited also tumor growth and progression in vivo. Vandana Relan1, Johanna Schagen2, Leanne Morrison3, Jennifer Martin3, Belinda E. Clarke4, Edwina E. Duhig3, Ian A. Yang5, Kwun M. Fong5, Conclusion: Together, these data suggest that the anti-tumorigenic Rayleen V. Bowman5 effect of KB9520 is mediated through ERβ and demonstrate that selective 1Uq Thoracic Research Centre,, University Of Queensland, The Prince targeting of ERβ may be an efficacious stand-alone treatment option of this Charles Hospital, Brisbane/QLD/AUSTRALIA, 2Uq Thoracic Research neoplasm and/or become an important add-on to existing therapy. Centre, University Of Queensland, The Prince Charles Hospital, QLD/ AUSTRALIA, 3The Prince Charles Hospital/AUSTRALIA, 4Department Disclosure: No significant relationships. Of Pathology, The Prince Charles Hospital, Brisbane/QLD/ AUSTRALIA, 5Department Of Thoracic Medicine, The Prince Charles Hospital, Brisbane/QLD/AUSTRALIA

POSTER SESSION 2 September 12, 2012 16:00-17:00 poster sessions | september 12 Background: Repurposing strategies offer potentially shorter times to implementation of novel therapy if drugs with anti-mesothelioma activity P2.13: EXPRESSION AND POST-TRANSLATIONAL MODIFICATIONS can be identified. Aim: We aimed to identify potentially active drugs by OF AKT ISOFORMS IN MALIGNANT PLEURAL MESOTHELIOMA screening a library of approved clinical compounds (Johns Hopkins Clinical CELLS Compound Library-JHCCL) for in vitro activity against a panel of human mesothelioma cell lines. Giulia Pinton1, Arcangela Gabriella Manente1, Ester Borroni1, Luciano Mutti2, Laura Moro1 Methods: Seven mesothelioma cell lines were screened against 1524 1Dept. Of Pharmaceutical Sciences, University Of Piemonte Orientale “A. JHCCL compounds using a growth inhibition assay with SYBR(R) Green I- Avogadro”, Novara/ITALY, 2Lab Of Clinical Oncology, Hospital Of Vercelli, fluorometric readout. Borgosesia (vc)/ITALY Results: At a final concentration of 10μM, 148 drugs produced at least 50% Background: The PI3K/AKT signaling pathway is aberrantly active and has growth inhibition on all seven cell lines. Compounds with a history of prior an important biologic impact in malignant pleural mesothelioma (MMe) cell oral or parenteral clinical use were retested in dose response experiments cycle progression and chemo-resistance. Akt consists of three isoforms, at final concentrations ranging from 100µM to 0.01µM. Seven drugs were Akt1, Akt2, and Akt3. Although these three isoforms are encoded by identified with IC50 (50% growth inhibitory concentrations) less than 1μM, separate genes, they share a common N-terminal PH domain, a catalytic six with IC50 1-5μM, and five drugs with IC50 5-10μM. Only five of the domain in the middle, and a C-terminus. The identity of the overall amino eighteen agents were known anti-neoplastic drugs. Other active agents acid sequence of the three isoforms is very high (~ 80%); however, the were previously approved as antibiotics, antihistamines, anti-helminthics, C-terminus and the PH-linker region are more diverse. Despite the growing anti-virals, anti-inflammatories, and immunomoulators. amount of research demonstrating the existence of isoform-specific regulation, many papers still draw generalized conclusions about Akt Conclusion: Active compounds were identified from a panel of function without considering the unique function of each Akt isoform. agents with history of clinical use. The anti-mesothelioma action Recent data have clearly demonstrated a role of SIRT1 in the modulation of several candidates in vitro now requires validation in vivo or in of AKT1 activation and a role of PARP1 as a gatekeeper for SIRT1 activity by clinical settings. This approach may yield improved treatments for limiting NAD+ availability. mesothelioma within a time frame responsive to predicted peak disease incidence. Acknowledgements: Cancer Australia, Dust Diseases Board, international mesothelioma interest group Methods: We have explored the expression and the balancing between The Prince Charles Hospital Foundation and Slater & Gordon Mesothelioma acetylation and phosphorylation status of AKT isoforms and their Travel Fellowship. Conflict of Interest: None interactors in human MMe cell lines in vitro. Disclosure: No significant relationships. Results: We firstly describe that MMe derived cell lines express both AKT1 and AKT3 isoforms. We demonstrate an inverse correlation between AKTs acetylation and phosphorylation modulated by PARP1/SIRT1 activation status in MMe cells. By immunoprecipitation experiments we evidence that POSTER SESSION 2 September 12, 2012 16:00-17:00 in basal conditions AKT1 is in part acetylated and in part phosphorylated and became highly phosphorylated and completely de-acetylated upon P2.15: AGONISTIC ANTI-CD40 ANTIBODY IS EFFECTIVE AGAINST PARP1 inhibition. Interestingly in our cancer-cell model, AKT1 activation POST-OPERATIVE TUMOUR OUTGROWTH IN A MURINE MODEL related to PARP1 inhibition, is unable to modulate pro-survival signals, OF MESOTHELIOMA because the downstream pathway is interrupted at the level of its effector mTOR. Conversely, SIRT1 inhibition or silencing result in a more Andrea Khong1, Matthew D. Brown1, Fahmi A. Mohamad2, Justin Vivian3, evident AKT1 acetylation. Interestingly SIRT1 inhibition results in increased Amanda Cleaver1, Bruce W.S. Robinson4, Andrew J. Currie1 acetylation of different AKT1 interactors. 1School Of Medicine And Pharmacology, University Of Western Australia/ AUSTRALIA, 2Universitas Indonesia/INDONESIA, 3Department Of Surgery, Conclusion: In conclusion our results clearly show how both PARP1 and University Of Western Australia/AUSTRALIA, 4School Of Medicine And SIRT1 affect critical cellular pathways involved in MMe progression and offer Pharmacology, University Of Western Australia, Perth/WA/AUSTRALIA a model of a regulatory inter-relationship between these proteins. These data could be helpful for designing new effective therapeutic strategies. Background: Local recurrence remains a major cause of death in patients with mesothelioma undergoing surgical resection. Metastases can also Disclosure: No significant relationships. occur, especially in longer term survivors. Agonistic anti-CD40 antibody is an immunotherapy that enhances immune priming of effector CD8 T cells by antigen presenting cells (APCs), leading to increased anti- tumour activity. We investigated the use of anti-CD40 antibody for the treatment of post-operative recurrence and metastasis, with regional lymphadenectomy, in a murine model of mesothelioma.

iMig2012.org • Abstract Book 105 Methods: Subcutaneous AB1-HA mesothelioma tumours were induced Conclusion: NGR-hTNF showed promising PFS and OS in this study. in the flank region of BALB/c mice. Tumours were completely or partially Based on these results, NGR-hTNF 0.8 μg/m2 q1w is currently tested debulked (75%) on day 16, with or without lymph node removal. On in a placebo-controlled phase II trial as a maintenance treatment in the day of surgery, animals that underwent complete surgical resection MPM pts who did not progress after 6 cycles of first-line chemotherapy were re-challenged with AB1-HA at the surgical site (local recurrence) (NCT01358084) and in a double-blind phase III trial evaluating best or the opposite flank (metastasis). Animals were treated with anti-CD40 investigator’s choice ± NGR-hTNF in relapsed MPM (NCT01098266). (FGK45) either systemically (partial debulk) or into the tumour bulk upon emergence (recurrence and metastases). Disclosure: A. Lambiase - MolMed - Employment C. Bordignon - MolMed - Employment Results: Anti-CD40 treatment slowed metastatic growth relative to untreated controls (p = 0.020) and improved survival from metastasis (78.57 ± 10.96% cure, H.R. 5.62 – 125.00). Anti-CD40 also retarded the growth of local recurrences (p = 0.004) and improved survival from recurrence (50.00 ± 17.68% cure, H.R. 1.18 - 4.81). In a partial debulk situation, anti-CD40 slowed outgrowth of residual tumour but did not improve survival. Removal of the tumour draining nodes did not impair efficacy (p>0.05). poster sessions | september 12 Conclusion: We show that anti-CD40 is effective at preventing local recurrence and metastasis and improving survival when combined with complete and partial surgical resection, an effect which was independent of the route of administration and lymph node removal. These findings argue for the usefulness of anti-CD40 antibody as an adjuvant immunotherapy to cancer surgery, particularly in cases where regional lymphadenectomy is indicated.

Disclosure: No significant relationships.

POSTER SESSION 2 September 12, 2012 16:00-17:00

P2.16: NGR-HTNF IN RELAPSED MALIGNANT PLEURAL MESOTHELIOMA (MPM)

Gilda Rossoni1, Vanesa Gregorc1, Antonio Lambiase2, Claudio Bordignon2 1Oncology, Istituto Scientifico San Raffaele, Milan/ITALY, 2Clinical Development, Molmed/ITALY

Background: NGR-hTNF exploits the asparagine-glycine-arginine (NGR) peptide for selectively targeting tumor necrosis factor (TNF) to cancer endothelial cells. Tumor hypervascularity is independent predictor of poor overall survival (OS) in MPM patients (pts). international mesothelioma interest group Methods: We report long-term results of a phase 2 trial that assessed NGR-hTNF in MPM pts with performance status (PS) ≤ 2 and radiologic progressive disease (PD) after a pemetrexed-based regimen. NGR-hTNF was given at 0.8 µg/m2 every 3 weeks (q3w) in 43 pts and weekly (q1w) in 14 pts. Primary endpoint was progression free survival (PFS), with restaging done q6w by MPM-modified RECIST criteria, while secondary aims included disease control (DC) rate and OS. We also tested the impact on outcome of neutrophil to lymphocyte ratio (NLR) at baseline (median 3; interquartile range 2-5). Median follow-up was 35.3 months.

Results: Baseline characteristics were (n=57): median age 57 years; M/F 35/22; PS 0/1-2 31/26; EORTC score good/poor 45/12. Median treatment free interval on prior therapy was 4.3 months. Only one drug-related grade ≥ 3 adverse events (AEs) was noted, common grade 1/2 AEs being transient chills (75%). No higher toxicity was reported using the q1w schedule. Median PFS was 2.8 months (95% CI 2.2-3.3). PFS was significantly improved in pts who experienced chills on treatment (hazard ratio, HR 0.44 p=.005). DC rate was 46% (95% CI 32-59; 26/57 pts; 1 partial response, 25 stable diseases) and was maintained for a median time of 4.7 months (95% CI 4.0-5.4). OS rates at 1, 2 and 3 years were 47%, 16% and 8%, respectively. Median OS was longer in pts with DC than in those with early PD (16.2 v 8.3 months, respectively; p=.01). According to schedule, 6-month PFS rates were 11% and 36% and 3-year OS rates were 4% and 19% for q3w and q1w, respectively. In pts with DC, median PFS were 4.4 and 9.1 months and median OS were 13.3 and 24.8 months for q3w and q1w, respectively. By multivariate analyses, a PS of 0 (p=.003) and a low baseline NLR (p=.004) were the only variables associated with improved OS. Median OS in pts stratified by NLR ≤ 2, 3 to 4, and ≥ 5 were 15.7, 10.5, and 4.2 months, respectively (p=.0003 for trend).

iMig2012.org • Abstract Book 106 Poster Session 2 Imaging and Staging September 12, 2012 16:00-17:00

POSTER SESSION 2 September 12, 2012 16:00-17:00 POSTER SESSION 2 September 12, 2012 16:00-17:00

P2.18: CT FINDINGS OF 17 CASES OF PRIMARY PERICARDIAL P2.19: CLINICAL STAGING OF MALIGNANT PLEURAL MESOTHELIOMA (PPM) MESOTHELIOMA WITH PET/CT, WHICH MAY BE A USEFUL TOOL FOR DETECTING TUMOR INFILTRATION INTO THE BIOPSY SITES 1 2 3 4 Katsuya Kato , Fumikazu Sakai , Yasuyuki Kurihara , Naohiko Inase , poster sessions | september 12 Hakutaro Miura5, Yukio Takeshima6, Kouki Inai6 Koji Kawaguchi, Tetsuo Taniguchi, Yoshinori Ishikawa, Takayuki Fukui, 1Radiology, Okayama University Hospital, Okayama/JAPAN, 2Radiology, Tetsuya Mizuno, Futoshi Ishiguro, Kohei Yokoi Saitama Medical University International Medical Center, Hidaka/ Thoracic Surgery, Nagoya University Graduate School Of Medicine, Nagoya/ JAPAN, 3Radiology, St. Marianna University. School Of Medicine, Kawasaki/ JAPAN JAPAN, 4Tokyo Medical And Dental University, Tokyo/JAPAN, 5Yokosuka General Hospital Uwamachi, Yokosuka/JAPAN, 6Pathology, Nstitute Of Background: The surgical outcome of malignant pleural mesothelioma Biomedical & Health Sciences Hiroshima University, Hiroshima/JAPAN (MPM) is far from satisfaction. One of the reasons for the poor outcome is suspected for difficulty of the correct staging, and as a result, it might lead Background: Primary pericardial mesothelioma (PPM) is an extremely rare to incomplete resection. The purpose of this study is to investigate the tumor accounting for approximately 0.7% of all malignant mesotheliomas. accuracy of preoperative positron emission tomography (PET)/computed Since primary pericardial mesothelioma is an extremely rare condition, tomography (CT) using fluorodeoxyglucose for MPM. there is no radiologic study focusing on CT manifestations of PPM based on a large patient group. The aim of the present study is to characterize CT Methods: Fifteen consecutive patients who underwent extrapleural features of 17 cases with PPM. pneumonectomy (EPP) for MPM during the last 5 years in our hospital were enrolled in this study. We re-evaluated the preoperative PET/CT and Methods: We collected clinical, imaging, and pathological data of 20 cases pathological results, and analyzed the accuracy of radiographic staging. of pericardial mesothelioma from 18 medical institutes in Japan. The data collection and study were supported by a research grant for the exploration Results: There were 12 males and 3 females, with a mean age of 62 of asbestos-related malignant tumor by the Ministry of Environment years. Induction chemotherapy was administered in 13 patients, 2 of Japan. Clinico-radiologic-pathologic (CRP) diagnosis was performed whom had downstaging. All patients had received PET/CT, which were by the consensus of the expert team (3 radiologists, 5 physicians, and performed a mean of 23 days (range 6-58) before EPP. From the PET/ 3 pathologists) in order to verify the eligibility of the diagnosis. For CT findings, tumor invasion to the lung was suspected in 9 patients, mesothelioma analysis, each case was categorized into either of the 5 diaphragmatic invasion in 6, chest wall invasion in 6, pericardial invasion subcategories: “definitely not” (definitely not PMP), “unlikely” (unlikely in 5, peritoneal invasion in 4, tumor infiltration into the biopsy sites in 8, PMP), “possible”(possible PMP), “probable” (probable PMP) and ”definite” and mediastinal lymph nodes metastases in 4. All patients underwent EPP

(definite PMP). The cases with questionable and/or atypical mesothelioma with an en-bloc resection of biopsy sites. One patient died a month after international mesothelioma interest group findings were especially discussed. We retrospectively analyzed imaging the operation because of acute respiratory distress syndrome. Pathological findings in cases with CRP agreement with PPM in order to characterize examination proved tumor invasion to the lung in 14 patients, diaphragm imaging features. Imaging protocols varied due to the retrospective nature in 12, chest wall in 10, pericardum in 7, biopsy sites in 8, and metastases of the study. Images were interpreted by the consensus of 3 experienced to the mediastinal lymph nodes in 8, respectively, while no patient with diagnostic radiologists. peritoneal invasion was observed. There were positive surgical margins in 7 patients; chest wall in 4, biopsy sites in 2, and pericardium in 1, Results: The results of CRP were as follows; three cases were “Definitely respectively. The sensitivity and specificity of the preoperative PET/CT not” because of originating from pleura. “Possible” are 4 cases and were 64% and 100% at the lung, 50% and 100% at the diaphragm, 50% “Probable” are 8 cases and “Definite” are 5 cases. We finally determined and 80% at the chest wall, 57% and 88% at the pericardium, 0% and 67% 17 cases of PPM on the basis of the CRP assessment of this original study at the peritoneum,100% and 100% at the biopsy sites, and 13% and 57% group. Histological sub-classification was as follows: 13 epithelial, 2 for mediastinal nodal metastases, each. All 5 patients who had complained biphasic, each of sarcomatoid and desmoplastic type. The imaging findings chest pain preoperatively were proven to have tumor invasion to the chest of the pericardium in 17 cases were as follows: no thickening, 4 cases wall. (24%); mild thickening, 3 (18%); irregular thickening, 6 (35%); and mass formation, 4 (24%), thus, approximately 41% of the cases showed no or Conclusion: MPM frequently invades to the biopsy sites, and PET/CT is mild thickening and were not indicative of malignant nature. Two cases demonstrated to have a high diagnostic accuracy for the tumor infiltration with mass formation mimicked mediastinal tumor. Pericardial thickness in this study. However, because it is difficult to evaluate the depth of ranged from 0 cm to 7.2 cm (median, 1.4cm). Pericardial effusion was invasion to chest wall and mediastinal lymph node metastases with PET/ observed in 13 of the 17 cases (76%); Pleural effusion was detected in CT, an intraoperative careful staging is indispensable. 12(70%), pleural effusion on bilateral, 8; right-sided, 2; and left-sided, 2. Six patients (35%) showed mediastinal lymph node swelling greater than 10 Disclosure: No significant relationships. mm in minimum diameter. Pleural plaque was found in 4 cases (24%).

Conclusion: The imaging findings of PPM had a considerably wide spectrum and included not only pericardial masses but also simple pericardial effusion and mediastinal mass formation. It is very important to know the imaging spectrum in order to make a precise diagnosis of PPM.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 107 POSTER SESSION 2 September 12, 2012 16:00-17:00 thoracic surgery patients the incidence was 4%. The incidence of VTE for malignant pleural mesothelioma (MPM) patients undergoing pleurectomy P2.20: DOES 18FDG PET PREDICT SURVIVAL IN MALIGNANT or extrapleural pneumonectomy (EPP) is unknown. VTE can worsen PLEURAL MESOTHELIOMA (MPM)? pulmonary hypertension, and significantly affect patient morbidity and mortality. Database analysis was performed to determine the incidence Astero Klabatsa1, Sugama Chicklore2, Loic Lang-Lazdunski1, Sally and associated risk factors for VTE. Barrington2, Gary Cook2 1Thoracic Surgery, Guy’s And St Thomas’ Nhs Foundation Trust, London/ Methods: A retrospective cohort study of patients with MPM who UNITED KINGDOM, 2Clinical Pet Centre, Division For Imaging Sciences, underwent pleurectomy or EPP from 2006 to 2010. The primary end point King’s College London, /UNITED KINGDOM was occurrence of VTE within 30 days postop or during hospitalization. DVT was diagnosed by ultrasonography and PE by CT angiography. We Background: Malignant pleural mesothelioma (MPM) is a malignancy in selected candidate variables based on established risk factors for VTE. which the prognostic information provided by 18FDG PET imaging could Univariate analysis was performed to determine the incidence of VTE be particularly valuable in determining whether to pursue aggressive in relationship to the candidate variables. We started with saturated treatment. In this retrospective study, we aimed to identify independent model including all interested variables, then performed model reduction predictors of survival related to imaging in MPM. (backward selection) by excluding variables from the model with a p-value of >0.10 based on log likelihood ratio test. Methods: We retrospectively analysed baseline 18FDG PET/CT scans in poster sessions | september 12 61 patients with MPM scanned between 2006 and 2011. All scans were Results: 266 patients were included in the analysis. All received performed before medical treatment or surgery, including talc pleurodesis. postoperative DVT prophylaxis with sc heparin. The incidence of VTE was 22.2%; DVT 20.6 % and PE 1.9 %. DVT occurred in lower extremity in17%, Volumes of interest were drawn around the lesion/s to measure SUVmax, in upper extremity in 6.4%, and in both upper & lower extremities in 2.6%. SUVpeak, SUVmean, and total lesion glycolysis (TLG). One patient was excluded as there was no identifiable disease on the images. For each of the Independent variables associated with increased risk of VTE in saturated four 18FDG PET parameters measured, cases were grouped to low and high multivariate model were: presence of preop history of (h/o) smoking, DVT, grade according to cut-off points identified by maximal chi-squared test. heart failure, asbestos exposure, duration of surgery, periop coagulopathy, Differences in overall survival (OS) were assessed by the Kaplan-Meier test. transfusion of FFP, need for vasopressors; postoperative development Independent samples t-test and COX regression were used for correlations of HIT & acute kidney injury (AKI). In the reduced model, variables that between different histological groups and the presence of independent remained after backward selection were: preop h/o smoking, DVT, heart prognostic factors within our dataset. failure, asbestos exposure; duration of surgery, postoperative development of HIT & AKI. The variables most strongly associated with VTE were: h/o Results: Follow up was available for a median of 11.5 months (range 2 preop DVT and postoperative HIT, AKI. to 60 months). All 18FDG PET parameters above the cut-off points (high- grade) were associated with poorer survival. The average SUVmax was 13.04, and overall survival (OS) for the high-grade group was 10 months vs. the low-grade group of 13 months (p=0.027). The average SUVmean was 3.66, and overall survival (OS) for the high-grade group was 11 months vs. the low-grade group of 14 months (p=0.022). Average TLG was 665, and overall survival (OS) for the high-grade group was 9.5 months vs. the low-grade group of 14.5 months (p=0.024). Average SUVpeak was 8.48 and OS was 10 months for high-grades vs.13.5 months for low-grades; however no statistical significance could be demonstrated (p=0.085). No differences were found in any of the 18FDG PET parameters between different histological subtypes. COX regression analysis identified age and epithelioid and biphasic histology subtypes as independent prognostic international mesothelioma interest group factors.

18 Conclusion: FDG uptake in MPM, as measured by SUVmax, SUVmean or TLG, has prognostic significance and may be a helpful factor in stratifying individual patient management.

Disclosure: No significant relationships.

Conclusion: Despite periop DVT prophylaxis with heparin, MPM patients POSTER SESSION 2 September 12, 2012 16:00-17:00 are at high risk for VTE. We found three modifiable risk factors: preop history of DVT, postoperative development of HIT, & AKI. Patients with VTE P2.21: PERIOPERATIVE RISK FACTORS FOR VENOUS had significantly delayed ICU and hospital discharge. Hence, modification THROMBOEMBOLISM IN PATIENTS WITH MALIGNANT PLEURAL of these risk factors should be considered by initiating more intense MESOTHELIOMA: A RETROSPECTIVE ANALYSIS OF OUR FOUR prophylaxis earlier. YEAR DATABASE. Disclosure: No significant relationships. Sujatha Pentakota1, Cindy Gonzalez2, David J. Sugarbaker3, Gyorgy Frendl1 1Anesthesiology, Perioperative, And Pain Medicine, Brigham And Women’s Hospital, Boston/MA/UNITED STATES OF AMERICA, 2Surgery, Brigham And Women’s Hospital, Boston/MA/UNITED STATES OF AMERICA, 3Division Of Thoracic Surgery, Brigham And Women’s Hospital, Boston/MA/UNITED STATES OF AMERICA

Background: Venous thromboembolism (VTE), including deep vein thrombosis (DVT) and pulmonary embolism (PE), increases morbidity and mortality in cancer patients. The analysis of 22 million cancer patients showed 3.5% incidence of VTE within 30 days postop, for

iMig2012.org • Abstract Book 108 POSTER SESSION 2 September 12, 2012 16:00-17:00

P2.22: REPRODUCIBILITY OF THERAPY RESPONSE EVALUATION BETWEEN EXPERIENCED AND LESS EXPERIENCED READERS OF PLEURAL MESOTHELIOMA BY MRECIST, RECIST 1.0, RECIST 1.1, AND WHO

Thorsten Persigehl1, Leandra Stahlhut1, Alexander C. Bunck1, Yongqiang Tan2, David Maintz1, Lawrence Schwartz2, Binsheng Zhao2 1Radiology, University Hospital Cologne, Cologne/GERMANY, 2Radiology, Columbia University Medical Center, New York/UNITED STATES OF AMERICA

Background: Tumor therapy monitoring of malignant pleural mesothelioma is difficult due to the specific tumor growth pattern. Today there exists a wide number of response evaluation criteria, such as modified RECIST (mRECIST; up to 2 target lesions on 3 anatomical levels), RECIST 1.0 (up to 5 poster sessions | september 12 target lesions), RECIST 1.1 (up to 2 target lesions), and WHO (up to 5 target lesions). The aim of this study was to evaluate the accordance of tumor size change measurements between experienced and less experienced readers for the above mentioned response evaluation criteria.

Methods: CT scans from 15 patients with a total of 43 baseline and follow- up examinations from a clinical multi-center pleural mesothelioma trial were retrospectively analyzed. One experienced radiologist (≥10 years) and one less experienced radiologist (≥1 year) selected target lesions independently according to mRECIST, RECIST 1.0, RECIST 1.1, and WHO criteria. CT scans were re-measured after an interval of greater than one month. The association of the relative changes in the measurements between baseline and follow-up(s) were analyzed for inter- and intra- reader variability using the different response criteria. Inter- and intra- reader variability was analyzed by Bland-Altman method.

Results: Our preliminary data from two radiologists demonstrated the best inter-reader accordance by using RECIST 1.0 within the 95% limits of agreement between the experienced reader and less experienced reader of -9.4/ 18.2. In contrast, intra-reader variability of the experienced reader was best using mRECIST within the 95% limits of agreement of -6.3/ 4.8. WHO performed worst with an inter- and intra-reader variability of -34.0/ 38.6 and -29.1/ 20.0, respectively.

Conclusion: In our study, pleural mesothelioma treatment response evaluation by an experienced and less experienced reader was most international mesothelioma interest group reliable when using RECIST 1.0, possibly due to depiction of better suitable target lesions. However, mRECIST demonstrated the best reproducibility with the experienced reader. Higher variability was seen with RECIST 1.1. WHO was consistently poorly suitable to evaluate tumor response. To confirm these findings, the inclusion of further readers and more patients’ data is warranted.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 109 Poster Session 2 Pathology and Cytology September 12, 2012 16:00-17:00

POSTER SESSION 2 September 12, 2012 16:00-17:00 histopathologic examination of an adequate thorascopic or open biopsy. Since DMM is often heterogeneous, a biopsy may not be representative of P2.23: PATTERNS OF LYMPH NODE SPREAD TO N2 NODES the entire tumor. The goal of this study was to determine the accuracy of PREDICTS SURVIVAL IN PATIENTS WITH BIPHASIC PLEURAL pretreatment biopsy in establishing the histopathologic type of DMM. MALIGNANT MESOTHELIOMA (MM)

Methods: We examined 151 consecutive patients with pleural DMM treated poster sessions | september 12 Lucian Chirieac1, William G. Richards2, David J. Sugarbaker3 from 1988 to 1997 at Brigham and Womens Hospital by extrapleural 1Department Of Pathology, Brigham And Women’s Hospital, Boston/ pneumonectomy (EPP) followed by heated chemotherapy all of whom had MA/UNITED STATES OF AMERICA, 2Division Of Thoracic Surgery, Brigham a pretreatment biopsy available for review. We characterized the presence And Women’s Hospital And Harvard Medical School, Boston/MA/UNITED of epithelioid and sarcomatoid histology in the resection and pretreatment STATES OF AMERICA, 3Division Of Thoracic Surgery, Brigham And Women’s biopsy specimens. Associations between the histology in pre- and post- Hospital, Boston/MA/UNITED STATES OF AMERICA treatment specimens were investigated.

Background: In patients with diffuse MM metastases to extrapleural N2 Results: The histology type of DMM in pretreatment biopsies were lymph nodes are a poor prognosis characteristic. Studies from our group epithelioid in 120 patients (79%), mixed in 21 patients (14%), sarcomatoid have shown that metastases to N2 lymph nodes from biphasic MM have in 8 patients (5%), and indeterminate in two patients (1%). The histology either both epithelioid and sarcomatoid histologies or only the epithelioid type of DMM in resection specimens was epithelioid in 93 patients (62%), histology, but the clinical significance of this observation is unknown. mixed in 51 patients (34%), and sarcomatoid in 7 patients (4%). Biopsy In this study we investigated the clinical significance of the component findings were concordant with resection findings in 116 patients (Spearman metastatic to N2 lymph nodes from patients with biphasic MM. r=0.64, p<0.0001).

Methods: We identified 231 consecutive patients with biphasic MM treated Conclusion: Our data suggests that a diagnosis of mixed or sarcomatoid by surgery at Brigham and Women’s Hospital between 1988 and 2009 and DMM in the pretreatment biopsy is highly predictive of the histology in the found 74 with metastases to mediastinal N2 lymph nodes. We evaluated resection specimen. A diagnosis of epithelioid DMM in the pretreatment the N2 lymph node metastases of 41 of these patients with biphasic MM biopsy is less accurate, and it changed to a less favorable one in a and available pathology material and correlated the findings with overall significant proportion of the cases. The results of our study emphasize survival. the importance of thorough biopsy sampling in patients with malignant mesothelioma and the value of resection specimens for accurate diagnosis. Results: All 41 patients (8 F/33 M; mean age 62; range 31-88) had a diagnosis of biphasic MM metastatic to N2 lymph nodes. Twenty-four Disclosure: No significant relationships. patients (59%) with biphasic MM had both epithelioid and sarcomatoid components in the N2 lymph nodes and seventeen patients (41%) showed international mesothelioma interest group spread only of the epithelioid component to the N2 lymph nodes. The POSTER SESSION 2 September 12, 2012 16:00-17:00 mean follow-up period after surgery was 11.2 months. The median survival of patients with mixed histology in the N2 lymph nodes was 8.9 months versus 11.9 months for those with an epithelioid component (p=0.059). P2.25: BINDING OF CA125 TO MESOTHELIN IN THE SERUM OF MALIGNANT MESOTHELIOMA PATIENTS Conclusion: Our data indicate that the presence of a mixed component 1 1 1 1 in the N2 lymph nodes may predict a worse overall survival in patients Jenette Creaney , Ian Dick , Hanne Dare , Yvonne Demelker , Arthur W. 2 3 with biphasic MM. The results of our study emphasize the importance Musk , Bruce W.S. Robinson 1 of histologic classification of not only the surgical specimen but also the School Of Medicine And Pharmacology, University Of Western Australia, 2 lymph node metastases and highlight the biologic complexity of disease Nedlands/AUSTRALIA, Respiratory Medicine, Sir Charles Gairdner Hospital, 3 progression in biphasic MM. Nedlands/WA/AUSTRALIA, School Of Medicine And Pharmacology, University Of Western Australia, Perth/WA/AUSTRALIA Disclosure: No significant relationships. Background: One promising biomarker for malignant mesothelioma (MM) is soluble mesothelin, which is elevated in the serum of half of MM patients at diagnosis and approximately 15% before radiological or clinical signs of POSTER SESSION 2 September 12, 2012 16:00-17:00 MM. Mesothelin, a 40 kDa glycoprotein constitutively expressed on the cell surface of mesothelial cells, has been demonstrated to bind to the ovarian P2.24: THE ACCURACY OF PRETREATMENT BIOPSY OF PLEURAL cancer biomarker CA125. It has been suggested that metastatic spread MALIGNANT MESOTHELIOMA IN PREDICTING HISTOPATHOLOGIC of CA125 expressing tumours can be facilitated by binding to mesothelin TYPE IN THE EXTRAPLEURAL PNEUMONECTOMY SPECIMEN expressing tissues in the pleura and peritoneum and therapeutic targeting of this interaction is currently being investigated. We hypothesised that Lucian Chirieac1, David J. Sugarbaker2, Paul Vanderlaan1 binding of soluble mesothelin and CA125 in the serum may prevent 1Department Of Pathology, Brigham And Women’s Hospital, Boston/MA/ accurate quantification of mesothelin levels, and it was of concern that the UNITED STATES OF AMERICA, 2Division Of Thoracic Surgery, Brigham And diagnostic accuracy of mesothelin for MM could be being impaired. Women’s Hospital, Boston/MA/UNITED STATES OF AMERICA Methods: An ELISA-based assay was developed to detect mesothelin Background: Pathologic classification of diffuse malignant mesothelioma bound to CA125. Serum was collected from 41 patients with MM, and (DMM) into epithelioid, sarcomatoid, and biphasic types is an important levels of mesothelin, CA125 and of bound mesothelin-CA125 were predictor of survival. The diagnosis of DMM is usually based on determined. To dissociate mesothelin bound to CA125 serum samples

iMig2012.org • Abstract Book 110 were incubated in a solution of sodium sodium dodecyl sulphate (SDS) and POSTER SESSION 2 September 12, 2012 16:00-17:00 diethylenetriaminepenta acetic acid (DTPA). P2.27: BRAIN AND SACRAL METASTASIS IN A PATIENT WITH Results: Serum mesothelin concentrations ranged from 1 to 100 nM, with MALIGNANT MESOTHELIOMA 29 of the 41 cases being mesothelin positive. Serum CA125 concentrations ranged from 4 to 3400 U/mL, with 18 of the 41 cases being CA125 positive. Lukas J. Lee1, Ya-fang Chen2, Mu-zon Wu3 CA125 was demonstrated to be bound to mesothelin in nine of the MM 1Division Of Environmental Health And Occupational Medicine, National patient sera. In each of these nine cases the CA125 concentration was Health Research Institutes, Zhunan, Miaoli County/TAIWAN, 2Department above 35 U/mL and the mesothelin concentration was above 4 nM. All Of Medical Imaging, National Taiwan University/TAIWAN, 3Department Of nine patients were male, with predominantly epithelial tumours. Median Pathology, National Taiwan University/TAIWAN survival for the group of patients with evidence of CA125 bound to mesothelin in the serum was 5 months, which was significantly shorter Background: Malignant mesothelioma (MM) is a rare cancer primarily than for the group which did not have CA125 bound to mesothelin (median caused by asbestos exposure. In Taiwan, the incidence of MM has been survival 13.5 months; p = 0.019). The addition of SDS and DTPA to serum increased in recent years, around 1.4 per million per year for males. samples disrupted the observed binding of mesothelin to CA125, but no Neurological complication related to MM is very uncommon. change in detectable mesothelin levels was observed. Methods: We reviewed all sixty-six MM registered in the cancer registry Conclusion: CA125 binds to mesothelin in the serum of some MM system of an university-affiliated hospital during the past three decades poster sessions | september 12 patients. Binding was only observed in patients with relatively high CA125 and identified a young male case with frontal brain and sacral metastasis. and mesothelin levels. Binding could be disrupted but this did not alter We presented the clinical history, images, and pathological findings, and the amount of mesothelin detected in clinical samples. At the range of proposed a hypothesis of the rare metastasis in MM. mesothelin concentrations observed in the majority of non-symptomatic asbestos exposed individuals binding of CA125 would not impair the Results: A 35-year-old man had initial presentation of dry cough and body diagnostic accuracy of the mesothelin biomarker. weight loss, and then was diagnosed as malignant pleural mesothelioma based on pathological findings of ultrasound-guided pleural biopsy in Disclosure: No significant relationships. December 2005. He was lost to follow-up until November 2006, when progressive dyspnea aggravated. Then he visited our oncologic clinic and had received thoracentesis every week. Three months later he developed acute hypercapnic respiratory failure. After intensive care, he received POSTER SESSION 2 September 12, 2012 16:00-17:00 a course of chemotherapy with cisplatin and pemetrexed (Alimta®). He developed bilateral leg numbness and the spinal MRI showed abnormal P2.26: EXPRESSION OF MELANOMA ANTIGEN-ENCODING signal and enhancement at lower sacrum and the iliac bone suggestive of GENE-1(MAGE-1) IN MESOTHELIOMA CELLS AND REACTIVE metastases. He had acute consciousness changes in July 2007. Head CT MESOTHELIAL CELLS. scan showed a 4.7cm heterogeneously enhancing mass in right frontal lobe abutting the pial brain surface and a smaller lesion at the posterior margin Hironori Katayama1, Masataka Tanno1, Masaru Hosone1, Zenya Naito2, of right thalamus, suggestive of brain metastases. There was abnormal Shotaro Maeda3, Naoyuki Yoshino4, Tomomi Hirata4 leptomeningeal enhancement along adjacent brain suggestive of CSF 1Pathology, Nippon Medical School Tama-Nagayama Hospital, Tokyo/ process. The patient expired soon after the diagnosis of distant metastasis, JAPAN, 2Pathology, Nippon Medical School, Tokyo/JAPAN, 3Pathology And at the age of 36. Cytology Center, Mitsubishi Chemical Medience, Tokyo/JAPAN, 4Surgery, Tama-Nagayama Hospital, Nippon Medical School, Tokyo/JAPAN Conclusion: Malignant mesothelioma is primarily caused by exposure to asbestos. Through a careful study on this unusual mesothelioma patient Background: Recently mesothelioma cases have been rapidly increased with both sacral spine and brain metastasis, we proposed a possible in number in Japan. Mesotheliomas occur in various forms in body cavity hypothesis of metastasis: CSF spread of malignant mesothelial cells, international mesothelioma interest group fluid. Immunohistochemical staining with a panel of antibodies is used breaking through some areas of defective blood brain barriers up to the to diagnose mesothelioma when the condition is strongly suggested arachnoid plexus to reach the right frontal brain, and in the CSF flow along by cytological findings. We performed Immunocytochemical staining a the spinal canal down to the sacral area. differential diagnosis of mesothelioma from reactive mesothelial cells using MAGE-1 antibody. Disclosure: No significant relationships.

Methods: Study subjects were 7 mesothelioma cases (pleural mesothelioma: 5 cases, peritoneal mesothelioma: 1 case, pericardial mesothelioma: 1 case) and 10 non-cancerous cases of reactive mesothelial POSTER SESSION 2 September 12, 2012 16:00-17:00 cells (pleural effusion: 7 cases, peritoneal effusion: 3 cases) evaluated at our department. Immunostaining was performed in cell block sections P2.28: HOW TO MAKE A DEFINITIVE DIAGNOSIS OF prepared from body fluid and also performed in cell transfer sections MESOTHELIOMA BY THE BODY FLUID CYTOLOGY. prepared from Papanicolaou-stained specimens. Three mesothelioma cases were also examined for expression in tissue. Immunostaining was Shotaro Maeda1, Hironori Katayama2, Masaru Hosone3, Tomomi Hirota4, performed with a Dako Autostainer and EnVision visualization system. Masataka Tanno4, Zenya Naito5 1Pathology And Cytology Center, Mitsubishi Chemical Medience, Results: 1) Positive results were obtained in 5 cases of mesothelioma Japan/JAPAN, 2Hironori Katayama/JAPAN, 3Nippom Medical School/ (70%), expressed in the nucleus. JAPAN, 4Nippon Medical School Tama-Nagayama Hospital/JAPAN, 5Nippon 2) Negative results were obtained in all cases(10 cases) of reactive Medical School/JAPAN mesothelial cells (100%). Background: In mesothelioma, body fluid accumulates in early stage (stage Conclusion: Using the Melanoma antigen-encoding gene (MAGE-1) I) different from other malignant tumors. Therefore, body fluid cytology antibody is effective in differentiating mesothelioma from reactive is very important for early detection and treatment of mesothelioma. mesothelial cells. Particularly, the prognosis of pleural mesothelioma in early stage has become better recently because of extra-pleural pneumonectomy or Disclosure: No significant relationships. pleurectomy/decortication. Therefore, misdiagnosis for mesothelioma in early stage by effusion cytology means that the survival chance of patient is lost. The purpose of this study is to determine whether malignant mesothelioma can be diagnosed definitively by effusion cytology using

iMig2012.org • Abstract Book 111 the cell transfer method and/or cell block method for immunochemical was defined if both p16INK4a/p14ARF signals were lost in nuclei and at least one staining. CEP-9 signal was observed. FISH was considered positive if homozygous deletion of p16INK4a/p14ARF locus was observed in at least >10% of cells which Methods: This study included 10 patients with MPM diagnosed by were identified histopathologically as mesothelial cells using adjacent effusion cytology in the past 6 years in our institute. These patients were hematoxylin & eosin-stained specimens. considered as having mesothelioma by body fluid cytology. Furthermore, immunochemical staining was carried out for a definitive diagnosis using Results: Homozygous deletion of the p16 INK4a/p14ARF locus was observed in the cell transfer method and/or the cell block method for these patients. atypical mesothelial cells invaded into subpleural adipose tissues. Atypical Tumor cells were immunostained using antibodies specific to calretinin, mesothelial cells on the seroral surface and in the submesothelial fibrous CK5/6, D2-40, mesothelin, thrombomodulin, CEA, MOC31, p53 protein, tissues of the same MPM patient also showed homozygous deletion of EMA, E-cadherin, CD146., MMP-9 and MAGE-1 . the p16 INK4a/p14ARF locus. In addition, homozygous deletion of the p16 INK4a/ p14ARF locus was observed in atypical mesothelial cells confined to the Results: serosal surface and the submesothelial fibrous tissues. Calretinin was positive in 10/10 patients (100%), CK5/6 in 9/9 (100%),D2- 40 in 10/10 (100%), mesothelin in 9/9(100%), thrombomodulin in 7/8 Conclusion: These results showing homozygous deletion of the p16INK4a/ (87.5%), CEA in 0/10 (0%), MOC31 in 0/8(0%), BerER4 in 0/8(0%), p53 p14ARF locus in atypical mesothelial cells on the serosal surface and in the protein in 9/9(100%), EMA in 8/9 (88.9%), E-cadherin in 7/8 (87.5%), submesothelial fibrous tissues indicate that FISH analysis of the p16INK4a/

CD146 in 6/7(85.7%), MMP-9 in 7/8(87.5%), MAGE-1 5/7(71.4%). As a p14ARF locus may be useful to identify early-stage MPM, such as genuine poster sessions | september 12 result, the 10 patients were definitively diagnosed as having MPM. p53 mesothelioma in situ. protein, EMA, E-cadherin, CD146, MMP-9 and MAGE-1 were especially useful for discrimination of mesothelioma and reactive mesothelial cells. Disclosure: No significant relationships. Histological examination and/or electron microscopical examination was finally done and the cytological diagnosis was confirmed.

Conclusion: Good immunochemical staining results were obtained, enabling the definitive diagnosis of mesothelioma using the cell transfer method and/or cell block method. Immunochemical staining using multiple antibodies is important for the definitive diagnosis of mesothelioma by cytology.

Disclosure: No significant relationships.

POSTER SESSION 2 September 12, 2012 16:00-17:00

P2.29: THE P16INK4A/P14ARF DELETION IN ATYPICAL MESOTHELIAL CELLS LOCALIZED ON THE SEROSAL SURFACE OR WITHIN THE SUBMESOTHELIAL FIBROUS TISSUES OF THE PARIETAL PLEURA

Tohru Tsujimura1, Ayuko Sato1, Ikuko Torii1, Misa Song1, Shinji Matsumoto2, Kazuki Nabeshima2, Seiki Hasegawa3, Takashi Nakano4 1Department Of Pathology, Hyogo College Of Medicine, Nishinomiya/ JAPAN, 2Department Of Pathology, Fukuoka University School Of Medicine international mesothelioma interest group And Hospital, Fukuoka/JAPAN, 3Department Of Thoracic Surgery, Hyogo College Of Medicine, Nishinomiya/JAPAN, 4Division Of Respiratory Medicine, Department Of Internal Medicine, Hyogo College Of Medicine, Nishinomiya/JAPAN

Background: Malignant pleural mesothelioma (MPM) is an aggressive tumor arising from mesothelial cells on the serosal surfaces of the pleural cavity. Since it is difficult to distinguish morphologically between genuine invasion of MPM and inflammation-induced infiltration of non-tumor mesothelial cells, the pathological diagnosis of MPM is not generally established until atypical mesothelial cells invade into subpleural adipose tissues. On the other hand, the p16INK4a/p14ARF (CDKN2A) locus at chromosome 9p21 has been reported to be frequently deleted in MPM, but not in non-tumor mesothelial cells, by fluorescence in situ hybridization (FISH) analysis. To establish the utility of FISH analysis of the p16INK4a/ p14ARF locus for the diagnosis of early-stage MPM, we examined whether the p16 INK4a/p14ARF locus is deleted in atypical mesothelial cells confined to the serosal surface or the submesothelial fibrous tissues of the parietal pleura.

Methods: Tissue specimens with atypical mesothelial cells invaded into subpleural adipose tissues were selected based on histopathological examination of the pleural tissues obtained by video-assisted thoracoscopic surgery. Tissue specimens with atypical mesothelial cells confined to the serosal surface and the submesothelial fibrous tissues were also selected. Dual-color FISH analysis was performed on formalin-fixed, paraffin-embedded sections using a Spectrum Green-labeled chromosome 9 centromeric probe (CEP-9) and a Spectrum Orange-labeled, locus specific p16INK4a/p14ARF probe (Abbott, Tokyo, Japan). Homozygous deletion

iMig2012.org • Abstract Book 112 Poster Session 2 Programmatic Approach and Educational Resources for Mesothelioma Care September 12, 2012 16:00-17:00

POSTER SESSION 2 September 12, 2012 16:00-17:00 ineligibility or refusal, study attrition, and discussion of satisfaction surveys will also be presented. Information from this pilot study could prove helpful P2.30: THE PSYCHOSOCIAL IMPACT OF MESOTHELIOMA AND in developing a way to connect these geographically-dispersed, disabled, THE PROMISE OF A VIRTUAL DISCUSSION GROUP FOR PATIENTS and distressed patients.

1 2 2 2 Elizabeth Blackler , Caraline M. Craig , Tatiana Starr , Maria Farberov , Disclosure: No significant relationships. poster sessions | september 12 Lee M. Krug3, Marjorie G. Zauderer4, Valerie Rusch5, Jimmie Holland2, R G. Key2 1Social Work, Memorial Sloan-Kettering Cancer Center, New York/NY/ UNITED STATES OF AMERICA, 2Psychiatry And Behavioral Sciences, POSTER SESSION 2 September 12, 2012 16:00-17:00 Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED STATES OF AMERICA, 3Department Of Medicine, Thoracic Oncology Service, P2.31: IMPROVING THE CARE OF PATIENTS WITH Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED STATES MESOTHELIOMA: DEVELOPMENT OF AN ONLINE EDUCATIONAL OF AMERICA, 4Department Of Medicine, Memorial Sloan-Kettering Cancer RESOURCE FOR HEALTH CARE PROFESSIONALS. Center, New York/NY/UNITED STATES OF AMERICA, 5Department Of Thoracic Surgery, Memorial Sloan-Kettering Cancer Center, New York/NY/ Liz Darlison1, Sally Moore2, Patricia Hunt2, Dale Russell2, David Gledhill2, UNITED STATES OF AMERICA David Brighton2 1Univeristy Hospitals Of Leicester, Mesothelioma Uk, Leicester/UNITED Background: Mesothelioma is often caused by asbestos traceable KINGDOM, 2Royal Marsden Nhs Trust, London/UNITED KINGDOM to a specific environmental or occupational exposure. This unique pathophysiology and identifiable responsibility can lead to blame and Background: This poster describes the development and delivery of anger directed at the person or organization that allowed the exposure to an interactive online educational resource: The Mesothelioma Care in occur. Few studies have examined the emotional burden of mesothelioma. Practice Module. This module aims to improve the knowledge and skills of Little is known about patients’ coping methods or interventions to improve health care professionals working with people affected by mesothelioma. their psychosocial symptoms. The resource demonstrates collaboration between clinicians and educators from Mesothelioma UK and The School of Cancer Nursing and Methods: The aim of this two-part study is to identify the psychosocial/ Rehabilitation at the Royal Marsden NHS Foundation Trust. The poster physical needs and quality of life (QOL) of a sample of patients with provides an overview of the content and delivery of the module. pleural mesothelioma and explore the feasibility and promise of an Internet-based discussion group for these patients. We are approaching Methods: The content of the module is delivered over a 12-week patients who have a diagnosis of pleural mesothelioma, are able to read/ programme with a two-week orientation period to familiarise students speak English, and are receiving care at Memorial Sloan-Kettering Cancer with the online learning environment and to each other prior to the start international mesothelioma interest group Center (MSKCC). Patients with significant cognitive dysfunction or severe of the module. The module has been designed using a variety of teaching psychiatric disturbance are excluded from the study. Part 1 consists of a methods to suit different individual learning styles. The module can be set of questionnaires assessing coping methods, interpersonal support, taken at degree or masters level, or as a stand-alone module. mood, anxiety, and overall QOL. Part 2 is a weekly, six session Internet- based discussion group facilitated by a social worker. Part 2 participants Results: Three modules have successfully run during 2010, 2011 and 2012. are assessed three times: at baseline, approximately 1 week post group Thirty eight students have participated in the modules, 32 nurses from sessions, and approximately 4 weeks post group sessions in addition to the UK, two from Australia, two from the USA, one from Japan and one a self-report satisfaction survey. Part 1 participants are not required to from South Africa. The poster also presents brief findings of student and participate in Part 2. We plan to recruit 90 participants for Part 1 and 30 facilitator evaluations of the module following the first two modules (2010 participants for Part 2. & 2011). Overall, evaluations are highly positive and suggest significant impact on students’ levels of skills, confidence, knowledge and ability to Results: Of 56 participants enrolled thus far in Part 1, 50 have completed support people affected by mesothelioma. The module is planned to run the questionnaires. Thirty-two were male and 18 were female. The median yearly. age is 65 years old (range 38-83). Forty-two participants self reported their race as White, 3 Black or African American, 1 Asian/Pacific Islander, Conclusion: We believe this module is the first of its kind in the UK 3 other, and 1 did not report. Seven participants identified themselves and perhaps in the World. The resource forms part of an over-arching as Spanish, Hispanic, or Latino. In baseline assessment of wellbeing strategy within the UK to enhance the quality of mesothelioma services using the FACT-L, participants on average indicated the most distress in and improve patient and family member experience of care in line with functional wellbeing, which includes the ability to work, enjoy life, accept recommendations in the Mesothelioma Framework (DH 2007). illness, and be content with current QOL (M = 16.80, SD = 5.34, Range = 4–28). Overall, participants showed the least distress in social well- Disclosure: No significant relationships. being, indicating feelings of closeness and high emotional support and communication with family and friends (M = 22.87, SD = 3.50, Range = 15–28). Fifteen of the 19 participants enrolled to Part 2 have completed participation. Three participants are pending start of the next group. Thus far, 11 of 15 participants reported that the group met or exceeded their expectations.

Conclusion: At study end we plan to describe the psychological and physical symptom burden and QOL of patients. Reasons for study

iMig2012.org • Abstract Book 113 POSTER SESSION 2 September 12, 2012 16:00-17:00 POSTER SESSION 2 September 12, 2012 16:00-17:00

P2.32: THE ROLE OF THE THORACIC NURSE CASE MANAGER P2.33: DEVELOPMENT OF AN EDUCATIONAL PROGRAM ABOUT (NCM) IN SUPPORTING A PATIENT THROUGH MULTIMODALITY NURSING CARE OF PATIENTS WITH PLEURAL MESOTHELIOMA THERAPY (MMT) FOR MESOTHELIOMA. IN JAPAN

Sophia F.D. Holden Yasuko S. Nagamatsu1, Yuji Natori2 Thoracic Surgery, Guys And St Thomas’ Nhs Trust, London/UNITED 1Global Health Nursing, St. Luke’s Ollege Of Nursing, Tokyo/JAPAN, 2Hirano KINGDOM Kameido Himawari Clinic, Tokyo/JAPAN

Background: A Case Study demonstrating the role of the Nurse Case Background: In 2010, 1,209 people died of mesothelioma in Japan Manager (NCM) in supporting patients undergoing multimodality (Ministry of Welfare, 2011). The number of deaths from mesothelioma therapy (MMT) for malignant pleural mesothelioma (MPM). Pleurectomy is rapidly growing and Murayama (2008) projected that number would Decortication (PD) is the preferred surgical option in patients suitable increase to 100,000 deaths in Japan from mesothelioma by the year for MMT in our unit. To be included; slides of patient numbers coming 2040. The rapid increase of patients with mesothelioma (MPM) created a to our unit with mesothelioma (100-150), the numbers suitable for demand for nursing care for mesothelioma that requires knowledge about MMT (approximately 15%). Undergoing MMT takes altogether about six such things as asbestos, mesothelioma and its treatments, and benefits. months and the role of the NCM is to support the patient throught the Unfortunately there were no textbooks with relevant knowledge or poster sessions | september 12 process; organising appointments, scans, multidisciplinary team referrals, educational programs for nurses about mesothelioma in Japan or Japanese. psychosocial and financial support, symtom control. On completion of Therefore, this study aimed to develop an educational program about MMT the NCM in partnership with the Surgeon supports the patient with nursing care of patients with pleural mesothelioma. referral for second line treatment, survivorship and palliative care. Methods: The program was systematically developed according to Methods: 48 year old gentleman diagnosed at Video Assisted Thorascopy the steps of Instructional Design (ID): needs assessment, goal setting, (VATS) biopsy in October 2010 in another unit, Epitheliod MPM. No other writing performance objectives, developing assessment tools, developing medical history. Referred to Guys Hospital: NCM Assessment process instructional strategy, and developing instructional materials. To motivate Holistic Needs Assessment (HNA), Pre-admission work up, bloods learners, the teaching concept of andragogy (adult-focused) was adopted. ECG, X-ray , scans, physical examination, psychological assessment, Furthermore, narratives by MPM patients were introduced to address care- social assessment, guided visit of intensive care and Thoracic units. givers attitudes. The Research Ethics Review Board, St. Luke’s College of Vats, repeat biopsy and talc November 2010 at Guys (for staging and Nursing (approval no. 11-034) approved this study. reaccumulated effusion). PET-CT done in December ruled out bulky disease, transdiaphragmatic growth and significant mediastinal Results: 1. Development of Program Twenty one nurses were interviewed invovement, N3 disease or distant metastases. Patient had a PS of 0, fit for to assess the needs of nurses caring for MPM patients. According to the total pleurectomy/decortication. PD with hyperthermic pleural perfusion needs of nurses, nine goals were set and knowledge, skills and attitude (povidone-iodine) January 2011 at Guys, post operative Nursing Care and were described as performance objectives. To achieve each performance the NCM involvement in this discussed. Patient discharged from Guys day objective, a variety of instructional methods such as lectures, group work, 7, discharge advice given by NCM pain control, exercise, wound care, diet role-playing and group discussion were taken according to the nature of / elimination and breathing exercises. Telephone follow up. Prophylactic the performance objectives. Finally the instruction about knowledge, skills, radiotherapy on thoracotomy and drains scars 21Gy in 3# February 2011 attitude content was reformed to fit within the units of the two-day main 6 cyles of adjuvanct chemotherapy, Alimta and Cisplatin March-May 2011 program (14.5 hours) with a follow-up program one month later (3 hours). Follow-up PET-CT 4 weeks after completion of chemotherapy and at one The units of program were shown in Table 1. Each program was set for year shows no recurrence / progression. An EORTC QLQ-C30 questionaire small groups up to 30 learners. All instructional materials were originally 18 months post MMT was carried out and a patient interview developed in Japanese. international mesothelioma interest group

Results: Included in the results throughout the case presentation will be slides of PET -CT images, reults of EORTC QLQ questionaire (overall Main Program health rated 4/7, overall quality of life rated 6/7) and samples of interview Day1 questions / answers. The role of the NCM is explored throughout the patient journey, which is focussed on best practice standards 1000-1040 Icebreaking recommended by the Mesothelioma Framework (2007) in supporting 1045-1135 Asbestos & Benefit patients. 1145-1235 Surgical treatment for MPM Conclusion: The NCM plays a vital role in supporting patients having 1335-1425 Chemotherapy & Radiotherapy for MPM MMT for mesothelioma. The case load of this specific patient group has increased, and with survival rates showing an improvement with 1435-1505 Narrative speech by patient with MPM advancements in treatment. This patient group is likely to continue to 1515-1605 Symptoms of MPM & Palliative care increase and role of the NCM in survivorship can be developed further. Currently the overlall five year survival is 33% and for patients with 1615-1700 Needs of patients and family according to the stage of epitheliod subtype with complete macroscopic resection is 46%, (most MPM recent data from this unit) Recommendations for further practice will be 1700-1800 Group discussion to use HNA or EORTC questionarres on diagnosis and at regular intervals Main Program throughout treatment and follow up and audit the results. Day2 Disclosure: No significant relationships. 0830-0920 Home visiting care & Care coordination 0930-1030 Assessment & Management of symptoms of MPM 1045-1245 Role play 1400-1430 Stress management for nurse 1430-1500 Post test Follow-up Program

iMig2012.org • Abstract Book 114 with pleural effusion suspected for Mesothelioma, in order to give them 1000-1130 Group discussion a proper diagnostic setting, a standardized therapeutic option and a 1130-1230 Care for MPM in U.S.A and U.K continuous palliative support. 1230-1300 Follow-up test Methods: Review of the Regional MPM Registry showed an increasing

incidence of this cancer in our high risk area; a multidisciplinary panel 2. Program Implementation was instituted to evaluate the efficiency of actual treatments and follow The four main programs followed by 10 follow-up programs were up, comparing our experience and results with literature. Pneumologists, conducted from October 2011 to February 2012. Completing the program Thoracic Surgeons, Oncologists, Radiotherapists, Radiologists, Nuclear were 110 of the original 188 nurses who applied, coming from 100 institutes Physicians, Palliativists and General Practitioners were divided into two in all areas of Japan. The effectiveness of the program will be shown in the groups to discuss and propose new strategies for the “diagnosis and presentation. staging” and for the “treatment and follow up”; the statistician structured Conclusion: The first educational program about Nursing Care of Patients a multistep pathway with quality indices. with Pleural Mesothelioma in Japan was developed and implemented. Results: After a 20 months comparison between local attitudes and Disclosure: No significant relationships. literature guidelines, the two groups proposed a Diagnostic and Therapeutic Pathway (PDTA) [figure 1] which includes: (1) first and

second stage investigations guided by a single case manager through poster sessions | september 12 multidisciplinary meetings, (2) multimodal treatment for stage I and II POSTER SESSION 2 September 12, 2012 16:00-17:00 epithelioid Mesothelioma (neoadjuvant chemotherapy, Pleurectomy and Decortication (P/D), adjuvant radiotherapy) and (3) territorial palliative P2.34: PROPOSAL FOR A LOCAL DIAGNOSTIC AND THERAPEUTIC care for advanced stage or unfit patients. For patients affected by stage III PATHWAY FOR MALIGNANT PLEURAL MESOTHELIOMA: THE epithelioid Mesothelioma we decided to plan a randomized trial comparing neoadjuvant chemotherapy followed by P/D versus chemotherapy and IMPORTANCE OF A MULTIDISCIPLINARY TEAM palliation (talc poudrage) to establish if these patients can benefit from surgery regarding their quality of life (measurement of pain and dyspnoea) Sara Tenconi1, Roberto Piro2, Debora Formisano3, Massimiliano Paci1, and, eventually, their survival. Non-epithelioid Mesothelioma patients and Cristian Rapicetta1, Giorgio Sgarbi1, Luigi Zucchi2, & Mesothelioma advanced stage eligible patients will be treated with chemotherapy and Multidisciplinary Team3 palliative radiotherapy in case of symptomatic localized disease. 1Cardio Thoracic Vascular Dept.Arcispedale Santa Maria Nuova, Istituto Di Ricovero E Cura A Carattere Scientifico, Thoracic Surgery Unit, Reggio 2 Conclusion: In literature there isn’t a consensus on the best therapeutic Emilia/ITALY, Cardio Thoraco Vascular Dept. Arcispedale Santa Maria options for MPM, due to very heterogeneous reported trials; nevertheless, Nuova, Istituto Di Ricovero E Cura A Carattere Scientifico, Pneumology, 3 International Societies and Cancer Networks have tried to licence Reggio Emilia/ITALY, Arcispedale Santa Maria Nuova, Istituto Di Ricovero guidelines. In releasing a PTDA we should take into account all these E Cura A Carattere Scientifico, Statistic And Clinical Epidemiology Unit, recommendations as well as the local settings; moreover, difficulties are Reggio Emilia/ITALY represented by the fact that MPM remains a rare disease and that we still don’t seem to have a “cure” for these patients. For this reason, we think Background: Malignant Pleural Mesothelioma (MPM) is an asbestos that all treatments should aim to improve survival and achieve symptoms correlated cancer which incidence is expected to increase in the next control, with the minor functional impairment. decade in Italy. Amongst international literature there isn’t agreement on management of these patients. At the IRCCS Hospital of Reggio Emilia we Disclosure: No significant relationships. decided to schedule a multidisciplinary pathway for patients who present international mesothelioma interest group

iMig2012.org • Abstract Book 115 Poster Session 3 Diagnostic and Predictive Biomarkers September 13, 2012 11:30-12:30

POSTER SESSION 3 September 13, 2012 11:30-12:30 POSTER SESSION 3 September 13, 2012 11:30-12:30

P3.01: DIAGNOSTIC POTENTIAL OF MICRORNAS (MIRS) IN P3.02: PTEN PROTEIN EXPRESSION PREDICTS SURVIVAL IN MALIGNANT PLEURAL MESOTHELIOMA (MPM) MULTIMODALITY TREATED MALIGNANT PLEURAL MESOTHELIOMA PATIENTS: A VALIDATION STUDY OF TISSUE MICROARRAYS 1 2 3 Morten Andersen , Morten Grauslund , Jesper Ravn , Jens Benn poster sessions | september 13 1 1 2 2 Sørensen4, Claus B. Andersen5, Eric Santoni-Rugiu6 Byron Bitanihirwe , Martina Friess , Svenja Thies , Lukas Frischknecht , 2 1 2 3 1Department Of Pathology, 5442, Rigshospitalet, Copenhagen Alex Soltermann , Mayura Meerang , Holger Moch , Marc De Perrot , 4 1 1 University Hospital, Copenhagen/DENMARK, 2Department Of Pathology, Ghassan Allo , Walter Weder , Isabelle Opitz 1 5432, Rigshospitalet, Copenhagen University Hospital, Copenhagen/ Division Of Thoracic Surgery, University Hospital Zurich, Zurich/ 2 DENMARK, 3Department Of Thoracic Surgery, 2151, Rigshospitalet, SWITZERLAND, Division Of Clinical Pathology, University Hospital 3 Copenhagen University Hospital, Copenhagen/DENMARK, 4Department Zurich, Zurich/SWITZERLAND, Division Of Thoracic Surgery, Toronto Of Oncology, 5074, Rigshospitalet, Copenhagen University Hospital, General Hospital And Princess Margaret Hospital, Toronto/ON/CANADA, 4 Copenhagen/DENMARK, 5Department Of Pathology, 5441, Rigshospitalet, Department Of Laboratory Medcine And Pathobiolog, Toronto General Copenhagen University Hospital, Copenhagen/DENMARK, 6Department Of Hospital And Princess Margaret Hospital, Toronto/ON/CANADA Pathology, Section 5432, Rigshospitalet, Copenhagen University Hospital, Copenhagen/DENMARK Background: Malignant pleural mesothelioma (MPM) is characterised by complex chromosomal aberrations, including chromosome 10 losses. Background: MPM is histologically difficult to distinguish from reactive The tumour suppressor gene phosphatase and tensin homologue (PTEN) mesothelial proliferations (RMPs), partly because proposed MPM-markers located on chromosome 10q23 plays an important role in different cancers are not specific, reproducible or validated enough (Zimling ZG, Jørgensen (Chalhoub et al., 2009). We previously assessed PTEN protein expression on A, Santoni-Rugiu E., Histopathology, In Press). MiRs are small non-coding a tissue microarray (TMA, n = 352) from historical samples, mainly derived RNA-strands (~22 nt) that post-transcriptionally regulate gene-expression from post-mortem examination with only limited clinical information and vital cellular processes. MiR-expression in other cancers has shown available (Opitz et al., 2008), and demonstrated a shorter overall survival diagnostic significance, but it is uncertain whether currently published (OAS) for patients with PTEN loss. The present study therefore aimed to miR-data may provide candidate biomarkers for differentiating MPM from evaluate the expression of PTEN in two prospectively collected patient RMP. To pursue this goal, we performed a miR-expression-screening in cohorts treated for the diagnosis of MPM. formalin-fixed paraffin-embedded diagnostic biopsies, surgically resected MPM-specimens, and corresponding surrounding non-neoplastic pleura Methods: The two independent cohorts of MPM patients were uniformly (NP). treated with 3 cycles of platinum-based induction chemotherapy (CTX) followed by extrapleural pneumonectomy (EPP) and optional adjuvant

Methods: We performed a Real Time(RT)-qPCR-based (Exiqon®) screening radiotherapy. A TMA was constructed that included 46 patients for cohort international mesothelioma interest group of 742 human miRs’ expression in preoperative biopsies, epithelioid 1 (2 - 4 cores) and 102 patients for cohort 2 (4 cores). Immunoreactivity MPM-, and NP-specimens from 5 patients treated with extra-pleural levels of cytoplasmic and nuclear PTEN were semi-quantitatively scored by pneumonectomy as part of trimodal protocol. The relevant identified different observers in a blinded fashion and expression was then correlated differentially expressed miRs were validated on tissue samples from 24 to clinical and pathological parameters, such as histological subtype, independent MPM-patients by RT-qPCR-TaqMan® MicroRNA Assays staging, OAS, and progression-free (PFS) survival.

(Applied Biosystems). Threshold-cycles (Ct) were determined with related SDS v1.4 software, comparatively analyzing PCR-data with a normalizer. Results: In contrast to our historical cohort which showed a 60% loss of Significant (p < 0.05) differences between independent/paired groups PTEN expression, PTEN was more intensely expressed particularly in the were detected by non-parametric Mann-Whitney/Wilcoxon matched-pairs cytoplasm of patients uniformly treated with induction CTX and EPP (i.e. tests. low cytoplasmic PTEN expression; 26% in cohort 1 and 28% in cohort 2). There was no significant correlation between PTEN expression and Results: We identified significant difference in expression of 4 cancer- the histological subtype. Cox regression survival analysis indicated that relevant miRs (down-regulation of miR-17-5p, -126 and -652 and increased cytoplasmic expression of PTEN was significantly associated with up-regulation of miR-221) that correctly differentiated MPM from RMPs longer PFS in both test cohorts (p=0.05; cohort 1 andp=0.01, cohort 2) and was not influenced by chemotherapy (comparable miR-expression- (Figure 1), but not with OAS. levels in surgical samples and diagnostic biopsies). We then tested if the 4 miRs could fulfill the International Mesothelioma Interest Group’s recommendations for a suitable MPM-marker (sensitivity/specificity > 80%) by generating receiver-operating-characteristics (ROC)-curves using the RT-qPCR-data. The IMIG-criteria were not met for miR-17-5p and -221, while miR-126 and -652 showed high sensitivity and specificity.

Conclusion: The cancer-relevant miR-126 and -652 have great potential as biomarkers for distinguishing MPM from RPMs, while miR-17-5p and -221, previously reported to be specific for MPM’s histological subtypes and to play a pathogenic role in MPM, are not entirely suitable for this differential diagnostic purpose. Further studies will explore miRs’ prognostic and predictive potential and fully validate by in situ-hybridization-techniques their possible use in histopathological diagnostics.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 116 between very low TS mRNA level (<= 5 percentile) and overall survival (OS, 31 vs. 18 months; P=0.042). Patients with very high ERCC1 mRNA levels (>=95 percentile) survived significantly longer (75 vs. 18 months; P=0.048). In a multivariable analysis by Cox proportional hazards model that controlled for histology, stage, gender, as well as TS and ERCC1 mRNA levels, completeness of resection in terms of macroscopic complete resection (MCR) remained the only significant prognostic factor (P<0.001).

Conclusion: Very low TS and very high ERCC1 mRNA levels might influence overall survival in patients with MPM undergoing multimodality treatment. MCR seems to be the most important prognosticator. However, the role of TS and ERCC1 in multimodality treated MPM patients is worth of prospective validation in larger multicenter trials.

Disclosure: No significant relationships. poster sessions | september 13 POSTER SESSION 3 September 13, 2012 11:30-12:30

P3.04: GLYCOPEPTIDE SERUM MARKERS FOR MALIGNANT PLEURAL MESOTHELIOMA DIAGNOSTICS

Ferdinando Cerciello1, Annalisa Nicastri2, Meena Choi3, Damaris Bausch-Fluck4, Annemarie Ziegler5, Olga Vitek6, Emanuela Felley- Bosco5, Rolf Stahel5, Ruedi Aebersold7, Bernd Wollscheid4 1Clinic Of Oncology, Laboratory Of Molecular Oncology And Institute Conclusion: We have confirmed the role of PTEN as a prognosticator Of Molecular Systems Biology, Department Of Biology, University 2 for survival in MPM patients as we have found a significant association Hospital Zuerich And Eth Zurich, Zuerich/SWITZERLAND, Department between high cytoplasmic PTEN protein expression and prolonged PFS in Of Experimental And Clinical Medicine, Magna Graecia University, 3 two different cohorts of MPM patients uniformly treated with induction Catanzaro/ITALY, Department Of Statistics, Purdue University, West 4 CTX and EPP. These results support our previous study implicating PTEN Lafayette, In/UNITED STATES OF AMERICA, Institute Of Molecular as a candidate tumor suppressor in MPM. Further study of this gene, and Systems Biology, Department Of Biology And Nccr Neuro Center For 5 correlation with the downstream Akt signaling axis, appears warranted. Proteomics, Eth Zurich, Zuerich/SWITZERLAND, Clinic Of Oncology, Laboratory Of Molecular Oncology, University Hospital Zuerich, Zuerich/ Disclosure: No significant relationships. SWITZERLAND, 6Department Of Statistics And Department Of Computer Sciences, Purdue University, West Lafayette, In/UNITED STATES OF AMERICA, 7Institute Of Molecular Systems Biology, Department Of Biology And Faculty Of Science, University Zurich, Eth Zurich And University Zurich, POSTER SESSION 3 September 13, 2012 11:30-12:30 Zuerich/SWITZERLAND

P3.03: THE ROLE OF THYMIDYLATE SYNTHASE AND EXCISION Background: Mass spectrometric (MS) techniques were used for the REPAIR CROSS-COMPLEMENTING GROUP-1 AS PREDICTORS identification of serum accessible candidate markers of malignant pleural international mesothelioma interest group OF SURVIVAL IN PATIENTS UNDERGOING MULTIMODALITY mesothelioma (MPM). After surfaceome analysis of MPM vs control cell lines to identify potential candidate markers of MPM, the clinical significance TREATMENT FOR MALIGNANT PLEURAL MESOTHELIOMA of selected candidate markers was verified in serum cohorts using multiplexed selected reaction monitoring (SRM). Servet Bölükbas1, Annette Fisseler-Eckhoff2, Melanie Demes3, Sonja Stallmann4, Michael Eberlein5, Joachim Schirren1 Methods: We used the cell surface capturing technology (CSC) for the 1Thoracic Surgery, Dr. Horst Schmidt Klinik, Wiesbaden/ quantitative investigation of the cell surface N-glycoproteins of four MPM GERMANY, 2Institutes For Pathology And Cytology, Dr. Horst Schmidt (2 epithelioid and 2 biphasic) and four control cell lines (pleura and lung Klinik, Wiesbaden/GERMANY, 3Institut Für Pathologie Und Zytologie, Hsk adenocarcinoma). SRM-assays were established for glycopeptides detected Wiesbaden, Wiesbaden/GERMANY, 4Pathology And Cytology, Dr. Horst in higher abundance in MPM and used for multiplexed SRM quantitative Schmidt Kliniken Wiesbaden/GERMANY, 5Division Of Pulmonary, Critical analysis in patient sera enriched for N-glycopeptides. Care And Occupational Medicine, Carver College Of Medicine, University Of Iowa, Iowa/UNITED STATES OF AMERICA Results: Surfaceome analysis revealed 500 N-glycopeptides, corresponding to more than 300 cell surface N-glycoproteins from MPM- Background: Most studies investigated the relationship between derived cell lines. 56 candidate biomarker peptides were selected for Thymidylate Synthase (TS) and Excision Repair Cross-Complementing verification and quantification in patients and donors serum cohorts using Group-1 (ERCC1) mRNA levels and outcome in inoperable patients with a single multiplexed SRM-assay. Cohorts consisted of age and sex matched malignant pleural mesothelioma (MPM). The purpose of the present study MPM and non-small cell lung cancer patients as well as healthy donors was to study the association between TS and ERCC1 and overall survival in (total number > 75). Regression modeling was applied to define peptide- patients undergoing multimodality treatment for MPM. based classifiers of MPM. Serum analysis of the mesothelioma marker soluble mesothelin–related protein (SMRP) confirmed the robustness of our Methods: Forty-one consecutive patients with histologically confirmed approach. with MPM previously treated with radical pleurectomy, adjuvant Cisplatin and Pemetrexed as well as iradiation of the intervention sites were Conclusion: The relative quantitative investigation of the MPM surfaceome retrospectively analyzed. TS and ERCC1 mRNA levels were evaluated by unveiled serum accessible MPM candidate biomarkers. SRM analysis of real-time polymerase chain reaction and correlated to prospectively serum cohorts revealed a multiplexed peptide-based classifier of MPM. documented data. Kaplan-Meier analyses, log–rank test and Cox regression analyses were used to estimate survival and to determine predictors of Disclosure: No significant relationships. survival.

Results: In the univariate analyses, there was a significant correlation

iMig2012.org • Abstract Book 117 POSTER SESSION 3 September 13, 2012 11:30-12:30 complementing group 1 (ERCC 1), ribonucleoside-diphosphate reductase subunit M1 (RRM1) and thymidylate synthase (TYMS) with the outcome of P3.05: CLINICAL AND MORPHOLOGICAL PROGNOSTIC FACTORS drug therapy of patients with mesothelioma is discussed controversial. IN PATIENTS WITH MALIGNANT PLEURA MESOTHELIOMA (MPM) According to Scarlatti and colleagues thymidylate synthase (TYMS) is a predictive factor for Pemetrexed response and the excision repair cross- Annette Fisseler-Eckhoff1, Heike Schröder2, Sonja Stallmann3, Servet complementing group 1 (ERCC1) seems to be predictive for Cisplatin Bölükbas4, Dirk Kaiser5, Monika Serke6 therapies. With this in mind we analysed the mRNA expression level of 1Institutes For Pathology And Cytology, Dr. Horst Schmidt Klinik, those DNA repair and synthesis genes in primary mesotheliomas and Wiesbaden/GERMANY, 2Clinic Södra Älvsborgs/SWEDEN, 3Pathology And corresponding recurrent tumours of 5 patients. Cytology, Dr. Horst Schmidt Kliniken Wiesbaden/GERMANY, 4Thoracic Surgery, Dr. Horst Schmidt Klinik, Wiesbaden/GERMANY, 5Thoracic Surgery, Methods: The tumours were classified and the mRNA expression level Helios Clinic Gmbh/GERMANY, 6Pneumology Iii, Lung Clinic Hemer/ was detected by real time polymerase chain reaction analysis. All Patients GERMANY received surgery and were treated with Cisplatin/Pemetrexed regimes.

Background: Malignant pleura mesotheliomas (MPM) are represented as Results: All 5 primary mesothelioma were classified as epitheliods. heterogeneous tumor group regarding their clinic-biological behavior and After therapeutic treatment all 5 patients suffered from recurrent histomorphological growth pattern. According to the maximum of applied mesotheliomas. The median survival after primary diagnosis is 15.41 industrial asbestos in the 70s and their latency period for decades, the months. High ERRC1 and RRM1 expression levels were detected in relapsed poster sessions | september 13 highest incidence occurs assumedly between 2010 and 2020. Therefore tumors as compared to primary mesotheliomas. In contrast low TYMS the identification of prognostic factors for treatment decisions of patients expression levels were found. with MPMs is urgent. Conclusion: According to our study, TYMS as a predictive factor cannot Methods: 191 cases of MPMs were analyzed concerning histological be confirmed. RRM1 and ERCC1 as DNA repair and synthesis genes seem subtypes in biopsies and corresponding surgical specimens. Prognostic to play an important role in relapsed mesothelioma after treatment with molecular markers like tumor suppressor genes p53, p63 and proliferation Pemetrexed/Platinum regimes. index Mib-1 as well as clinical parameters like TNM stage were correlated with overall survival of the patients. Disclosure: No significant relationships.

Results: The median age of men (n=165) was 62 years, the median age of women (n=26) 63.7. 77% epithelial (n=147), 6.8% sarcomatoid (n=13) and 16.2% biphasic (n=31) subtypes were classified. The reclassification of the POSTER SESSION 3 September 13, 2012 11:30-12:30 diagnosis in biopsies and surgical specimens demonstrated accordance in nearly 100% of epithelial subtypes, 60% of sarcomatoid subtypes and P3.07: ASSOCIATION BETWEEN (GT)N REPEATS POLYMORPHISM 37.5% of biphasic subtypes. Survivial of patients with epithelial subtype IN THE HEME OXYGENASE-1 GENE PROMOTOR AND THE was significantly prolonged as compared to survival of patients with SUSCEPTIBILITY TO MALIGNANT MESOTHELIOMA IN THE sarcomatoid and biphasic subtypes, respectively (14.5 vs. 7.5 and 9.9 JAPANESE POPULATION WITH ASBESTOS-EXPOSURE respectively months median survival time, p= 0.01, p= 0.012). 5% and 36% of MPMs were negative for p53 and p63. Proliferation indices were Kazuya Fukuoka1, Yoshihiro Fujimori2, Yoshie Yoshikawa3, Shusai under 10% in 33% of patients, between 10 and 50% in 48% of patients Yamada4, Taiichiro Otsuki4, Asuka Okada4, Kunihiro Tamura4, Chiharu and between 51 and 80 % in 19% of patients. At time of diagnosis mainly Tabata4, Tomoko Hashimoto-Tamaoki3, Takashi Nakano4 advanced tumor stages were found (T2, T3) with lymph node metastases 1Cancer Center, Hyogo College Of Medicine, Nishinomiya/JAPAN, 2Division in 22% of patients and distant metastases in 6%. Patients with negative Of Blood Transfusion, Hyogo College Of Medicine, Nishinomiya/ nodal status survived significantly longer as compared to patients with JAPAN, 3Genetics, Hyogo College Of Medicine, Nishinomiya/ lymph node metastases (p=0.004). JAPAN, 4Division Of Respiratory Medicine, Department Of Internal international mesothelioma interest group Medicine, Hyogo College Of Medicine, Nishinomiya/JAPAN Conclusion: As epithelial subtype is correlated with a significant better survival in patients with MPM a correct diagnosis by intensive Background: Malignant mesothelioma (MM) is an aggressive tumor that morphological investigation of biopsies and surgical specimens by arises from mesothelial cells lining the pleural or peritoneal cavity. The experienced pathologist is essential for patients’ prognosis. In patients with development of MM is closely associated with asbestos exposure which epithelial subtype lymph node metastases is a significant prognostic factor. induces oxidative stress. Heme oxygenase (HO)-1, a rate-limiting enzyme Significant correlations of cT- or pT-stages or M-stages with overall survival of heme degradation, plays a protective role against oxidative stress. The could not be demonstrated. Under the aspect of new therapeutic regimens HO-1 gene promoter carries (GT)n repeats whose number is inversely the establishment of further e.g. molecular prognostic markers is essential. related to transcriptional activity of the HO-1 gene. Although asbestos is closely associated with the risk of MM, there are some populations that do Disclosure: No significant relationships. not develop MM after exposure to asbestos. In this study, we hypothesized that HO-1 gene variations may influence the susceptibility to MM.

Methods: To investigate the relationship between the length POSTER SESSION 3 September 13, 2012 11:30-12:30 polymorphism of (GT)n repeats and the susceptibility to MM, we analyzed the HO-1 gene promoter in 44 asbestos-exposed subjects without MM and P3.06: ERCC1, RRM1 AND TYMS EXPRESSION LEVELS IN PATIENTS 78 asbestos-exposed patients with MM using polymerase chain reaction WITH RECURRENT MALIGNANT PLEURA MESOTHELIOMA (PCR)-based genotyping.

Annette Fisseler-Eckhoff1, Sonja Stallmann2, Servet Bölükbas3, Joachim Results: The number of repeats ranged from 16 to 38 with two peaks at Schirren3, Melanie Demes4 23 and 30 repeats. Polymorphism of (GT)n repeats were grouped into two 1Institutes For Pathology And Cytology, Dr. Horst Schmidt Klinik, classes of alleles: short (S) (n<24) and long (L) (≥ 24) and three genotypes: Wiesbaden/GERMANY, 2Pathology And Cytology, Dr. Horst Schmidt L/L, L/S and S/S. The proportions of allele frequencies in class L as well as Kliniken Wiesbaden/GERMANY, 3Thoracic Surgery, Dr. Horst Schmidt genotypic frequencies of L allele carriers (L/L, L/S) were significantly higher Klinik, Wiesbaden/GERMANY, 4Institut Für Pathologie Und Zytologie, Hsk in the asbestos-exposed patients with MM than in the asbestos-exposed Wiesbaden, Wiesbaden/GERMANY subjects without MM.

Background: Pemetrexed/Platinum agents are applied as first-line Conclusion: The long size of (GT)n repeat in the HO-1 gene promoter treatment for mesothelioma. The association of excision repair cross- is associated with a higher risk of MM in the Japanese population

iMig2012.org • Abstract Book 118 with asbestos-exposure. This is the first report to demonstrate that 1Laboratory Medicine, Karolinska Instiute, Stockholm/SWEDEN, 2Science the polymorphism in the HO-1 gene promoter is associated with the For Life Laboratory, Karolinska Institute/SWEDEN, 3Department Of Chest susceptibility to MM. Diseases, Eskisehir Osmangazi University Medical Faculty/TURKEY

Disclosure: No significant relationships. Background: Diagnosis of malignant mesothelioma relies on morphological examination of cells and is often conclusive first when the disease has reached an advanced stage. The additional measure of diagnostic biomarkers in effusions, serum or plasma has not been extensively adapted POSTER SESSION 3 September 13, 2012 11:30-12:30 by the clinical routine; a gap that is partly due to the lack of predictability of proposed and tested biomarkers. However, soluble biomarkers are still P3.08: NOVEL PLASMA PROTEINS ASSOCIATED WITH a compelling, noninvasive adjunct to today’s standard diagnostics, which PROGNOSIS IN MALIGNANT PLEURAL MESOTHELIOMA could speed up and strengthen diagnoses. Using mass-spectrometry to screen pleural effusions from patients with mesothelioma, lung cancer and Steven C. Kao1, Glen Reid1, Ardeshir Amirkhani2, Dana Pascovici2, mesotheliosis, we aim to discover new biomarkers to add diagnostic value Xiaomin Song2, Rozelle Harvie3, Nick Pavlakis3, Stephen Clarke3, Mark P. and improve an already studied biomarker panel. Molloy2, Nico Van Zandwijk1 1Asbestos Diseases Research Institute, Concord/NSW/ Methods: Pleural effusions from 6 mesothelioma patients, 6 lung cancer AUSTRALIA, 2Australian Proteome Analysis Facility, Sydney/NSW/ patients and 3+4 (2 pools) patients suffering from benign mesotheliosis poster sessions | september 13 AUSTRALIA, 3Medical Oncology, Royal North Shore Hospital, St Leonards/ were selected for this study. To decrease variance and increase proteome NSW/AUSTRALIA coverage the effusions were depleted of high abundant proteins (MARS- 14 affinity purification, Agilent®), trypsinized, mass tagged (iTRAQ™) Background: The search for novel biomarkers to define more successful and subsequently pooled as well as fractionated (ultra narrow isoelectric and individual treatment approaches represent an important challenge for focusing and nano-liquid RP-HPLC) before screened with the LTQ Orbitrap those involved in the care for patients with malignant pleural mesothelioma Velos (Thermo Fischer Scientific, San Jose, CA, USA). Analyses of the results (MPM). In this exploratory study, we have made a systematic approach to were done using Significant Analysis of Microarrays (SAM v. 3.11; Stanford investigate the proteins present in plasma of MPM patients and to associate Tools, univariate), SIMCA (Umetrics®, multivariate: Principal Component their level with the outcome of disease. Analysis; PCA and Partial Least Squares; PLS) and Ingenuity Pathway Analysis (IPA; Ingenuity®, network analysis). Methods: Plasma samples from twelve MPM patients (6 ‘short-’ and 6 ‘long-term’ survivors from parallel phase II studies investigating Results: More than 1300 proteins were identified in the screened patients thalidomide) were used for proteomic analyses. Our series included (FDR<5%). Univariate analyses (SAM) identified proteins with high samples from patients with epithelial and biphasic MPM (8 epithelial; 3 respectively low expression in effusions from mesothelioma patients. The biphasic subtype; and 1 undetermined). Plasma samples were immuno- findings include a number of known mesothelioma biomarkers (mesothelin, depleted of the 14 most abundant proteins prior to labelling for isobaric osteopontin and apolipoprotein-CI) as well as several novel candidates; tag for relative and absolute quantitation (iTRAQ) analysis using mass though, with rather high FDR values. Multivariate analysis and modeling spectrometry. The most promising candidates were chosen for selected (PCA and PLS) identified thrombospondin-1 and galectin-1. Validation reaction monitoring mass spectroscopy (SRM-MS) confirmation. Statistical has been initiated on a larger number of patient effusions. Preliminary analyses using T-Test of peak areas were used to identify proteins that were results indicate that high levels of superoxide dismutase-2 add diagnostic differentially expressed between the short- and long-term survivor groups. information for a mesothelioma while galectin-1 is indicative of lung cancer. Galectin-1 and apolipoprotein-CI might be prognostic markers in Results: Median survival of short- and long-term survivors (1.2 and 38.3 mesothelioma. Furthermore, thrombospondin-1 is elevated in patients months, respectively) differed significantly (p = 0.001). This was also the with asbestos pleuritis and is a potential early marker for mesothelioma. case for the neutrophil-to-lymphocyte ratio (NLR) that was significantly Network analysis (IPA) gives further information about this disease and the higher in the group of short-term surviviors (p = 0.03). Other baseline system we work with. international mesothelioma interest group characteristics did not reveal major differences between the short- and long-term survivors. The total number of proteins identified was 226 Conclusion: In this study, proteomic screening of pleural effusions have (1% false discovery rate [FDR]). A number of those were found to be provided candidates for diagnostic and prognostic use for malignant differentially expressed between short- and long-term survivors (≥1.2-fold mesothelioma. The validation of biomarker candidates will be presented change; p ≤ 0.05) by iTRAQ: selenoprotein P; tetranectin (TETN); insulin- as well as network information about mesothelioma shedding light on like growth factor-binding protein 2 (IBP2); osteonectin (SPARC); platelet pathophysiological aspects. basic protein (CXCL7); and attractin (ATRN). SRM-MS analysis revealed that the concentrations of attractin (p = 0.01), tetranectin (p < 0.001) and Disclosure: No significant relationships. selenoprotein P (p < 0.001) were higher in long-term survivors. In contrast, an increase in the concentration of IBP2 (p = 0.07) and CXCL7 (p = 0.01) correlated with shorter survival. POSTER SESSION 3 September 13, 2012 11:30-12:30 Conclusion: We have demonstrated the feasibility of using the iTRAQ and SRM-MS proteomic techniques to investigate potential prognostic P3.10: HIGH BLOOD PLATELET-TO-LYMPHOCYTE RATIO protein markers in plasma of MPM patients. The candidates identified will IS AN INDICATOR OF POOR PROGNOSIS IN MALIGNANT be further examined by validation experiments with Western Blot and/or PLEURAL MESOTHELIOMA UNDERGOING EXTRAPLEURAL ELISA, and by testing samples from a larger patient series. PNEUMONECTOMY

Disclosure: No significant relationships. Tetsuzo Tagawa1, Licun Wu2, Zhihong Yun2, Katrina Rey- Mcintyre2, Ronald Feld1, Natasha Leighl1, John Cho1, Shaf Keshavjee1, Marc De Perrot1 1Toronto General Hospital And Princess Margaret Hospital, Toronto/ON/ POSTER SESSION 3 September 13, 2012 11:30-12:30 CANADA, 2Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, University Of Toronto, Toronto/ON/CANADA P3.09: SEARCH FOR NEW MESOTHELIOMA BIOMARKERS: PROTEOMIC SCREENING OF PLEURAL EFFUSIONS Background: Asbestos induced chronic inflammation plays a key role in the pathogenesis of malignant pleural mesothelioma (MPM). Recently, several 1 2 2 3 Filip Mundt , Henrik Johansson , Jenny Forshed , Sertac Arslan , biomarkers which are related to systemic inflammation including platelet- 3 1 2 1 Muzaffer Metintas , Katalin Dobra , Janne Lehtiö , Anders Hjerpe to-lymphocyte ratio (PLR) and neutrophil-to-lymphocyte ratio (NLR) have

iMig2012.org • Abstract Book 119 been reported as independent prognostic markers of patients with MPM. MPM (n=97) Non-malignant (n=23) Mann-Whitney Our aim is to investigate the prognostic value of PLR and NLR for patients with MPM undergoing extrapleural pneumonectomy (EPP). Gene mean 0.49 0.48 P=1.00 X Methods: Of 85 patients who underwent EPP for MPM during January 2001 median 0.39 0.36 to April 2011 at Toronto General Hospital, 65 patients whose blood test results before initial therapy were available were retrospectively analyzed. THBS- mean 0.11 0.02 P<0.01 Clinicopathological factors including PLR and NLR were analyzed in relation 2 to survival. median 0.10 0.00

Results: Demographics are as follows; median age: 61 (range 21-71 years), Conclusion: The serum antibody-titer against THBS-2 can be a useful non- sex: 55 males and 10 females, histology: 48 epithelial, 12 biphasic, 3 invasive marker in the diagnosis of MPM. sarcomatoid and 2 others, treatment: 34 chemotherapy (CH) + EPP + radiotherapy (RT), 12 CH + EPP, 11 RT + EPP, 6 EPP + RT and 2 EPP alone. Disclosure: No significant relationships. Complete resection rate was 81.5%. Median survival time was 23 months and 3 year survival rate was 39.6%. In univariate analyses, female gender (p = 0.013), PLR < 300 (p = 0.013), absolute platelet count < 400 x 109/liter (p = 0.036) and pN0-1 (p = 0.044) were predictors of good POSTER SESSION 3 September 13, 2012 11:30-12:30 poster sessions | september 13 survival. Histology (epithelial, p = 0.051) and NLR < 4 (p = 0.053) were not associated with overall survival. Multivariate analyses confirmed that P3.12: SERUM THIOREDOXIN-1 AND HISTONE DEACETYLASE female gender (p = 0.012) and PLR < 300 (p = 0.015) as an independent ACTIVITY IN PERIPHERAL BLOOD OF PATIENTS WITH MALIGNANT predictors of overall survival. PLEURAL MESOTHELIOMA Conclusion: High Platelet-to-lymphocyte ratio was independently Takashi Nakano, Kunihiro Tamura, Shingo Kanemura, Eisuke Shibata, associated with poor prognosis in patients with MPM undergoing EPP. Hisaya Ohkuwa, Taiichiro Otsuki, Hitomi Kamiya, Miki Honda, Koji Mikami, Yoshitaka Nogi, Risa Maeda, Takayuki Terada, Asuka Okada, Disclosure: No significant relationships. Shusai Yamada, Kazuya Fukuoka, Chiharu Tabata, Kozo Kuribayashi Division Of Respiratory Medicine, Department Of Internal Medicine, Hyogo College Of Medicine, Nishinomiya/JAPAN POSTER SESSION 3 September 13, 2012 11:30-12:30 Background: Asbestos-initiated oxidative stress significantly contributes P3.11: NOVEL MALIGNANT-MESOTHELIOMA-ASSOCIATED to carcinogenesis in asbestos-related malignancies by promoting oxidative DNA damage and regulating redox signaling pathways. Thioredoxin-1(TRX) ANTIGENS (GENE-X AND THBS-2) IN THE DIAGNOSIS OF is a small redox-active protein that possesses antioxidative activity and MALIGNANT PLEURAL MESOTHELIOMA (MPM) acts as a redox-regulating multifunctional protein. Oxidative stress also influences histone deacetylase (HDAC) activity which modulates gene 1 1 1 Masaru Takenaka , Fumihiro Tanaka , Yoshiki Shigematsu , Hidetaka expression through the deacetylation of lysine residues on histone proteins 1 1 2 2 Uramoto , Takeshi Hanagiri , Kazue Yoneda , Masaki Hashimoto , and acts transcriptional repressors of genes. Teruhisa Takuwa2, Seiji Matsumoto2, Yoshitomo Okumura2, Nobuyuki 2 3 4 2 Kondo , Kazuya Fukuoka , Takashi Nakano , Seiki Hasegawa Methods: We evaluated TRX in serum and HDAC activity in the peripheral 1 Second Department Of Surgery, University Of Occupational And blood mononuclear cells (1×105) of patients with malignant pleural 2 Environmental Health, Kitakyusyu/JAPAN, Thoracic Surgery, Hyogo mesothelioma (MPM). College Of Medicine, Nishinomiya/JAPAN, 3Division Of Respiratory international mesothelioma interest group Medicine, Department Of Internal Medicine, Hyogo College Of Medicine, Results: The patients with MPM had significantly higher levels of TRX in 4 Nishinomiya/JAPAN, Respiratory Medicine, Hyogo College Of Medicine, serum and higher HDAC activities than control population. MPM patients Nishinomiya/JAPAN with early clinical stage showed an increase in HDAC activity, and there was no significant difference among the IMIG clinical stages. On the other Background: Malignant pleural mesothelioma (MPM) is a highly aggressive hand, the patients with advanced stage MPM showed higher levels of TRX malignant tumor of the pleura associated with asbestos exposure, than those with early stage MPM. and its diagnosis is usually difficult at early stage. We identified novel mesothelioma-related antigens, Gene-X and thrombospondin-2 (THBS- Conclusion: MPM patients had a markedly increased level of TRX in serum 2), that were recognized by tumor-infiltrating B cells, and preliminary and high HDAC activity in peripheral blood mononuclear cells. These study demonstrated that these antigens were potentially useful diagnostic findings suggest that TRX and HDAC activity can be biomarkers in patients marker in MPM (Cancer Sci 2009). with MPM.

Methods: A total of 120 patients, who presented with a suspicion of Disclosure: No significant relationships. MPM and received pleural biopsy, were reviewed; 97 patients were finally diagnosed with MPM and 27 were with non-malignant diseases (NM). The antibody-titers against Gene-X and THBS-2 in the sera were measured by ELISA method.

Results: The serum antibody-titer against THBS-2 was significantly higher in MPM patients than in NM (P<0.01), but there was no difference in the serum antibody-titer against Gene-X (Table). The receiver operating characteristic (ROC) curve analysis showed a significant diagnostic value of serum antibody-titer against THBS-2 with the area-under curve of 0.886 (95% CI, 0.797 - 0.975; P<0.001) in discrimination of MPM from NM diseases; the sensitivity and specificity, when the cut-off value was 0.08, were 72.2% and 95.5%, respectively. There was no significant difference in serum antibody-titer against THBS-2 according to histologic subtype or clinical stage.

iMig2012.org • Abstract Book 120 Poster Session 3 Multimodality September 13, 2012 11:30-12:30

POSTER SESSION 3 September 13, 2012 11:30-12:30 stages were similar to other series, complete microscopic resection or response to chemotherapy were common in long term survivors. There P3.13: LONG-TERM (>2 YEARS) MALIGNANT PLEURAL appears to be no difference in long term survival in terms of treatment MESOTHELIOMA SURVIVORS FOLLOWING CHEMOTHERAPY AND modality, if patients survive longer than 2 years. MULTIMODALITY TREATMENT HAVE SIMILAR FEATURES AND

Disclosure: No significant relationships. poster sessions | september 13 SURVIVAL OUTCOMES

Hasan F. Batirel1, Guntulu Ak2, Muzaffer Metintas2, Onur Ermerak1, Fulden Yumuk3, Hale Basak Ozkok4, Volkan Kara1 POSTER SESSION 3 September 13, 2012 11:30-12:30 1Thoracic Surgery, Marmara University Hospital, Istanbul/ TURKEY, 2Department Of Pulmonary Medicine, Osmangazi University P3.14: SARCOMATOID PREDOMINANT MALIGNANT PLEURAL Hospital, Istanbul/TURKEY, 3Division Of Medical Oncology, Department Of Internal Medicine, Marmara University Hospital, Istanbul/ MESOTHELIOMA: AN INSTITUTIONAL APPROACH AND TURKEY, 4Department Of Radiation Oncology, Acibadem University, EXPERIENCE Istanbul/TURKEY Robert B. Cameron1, Olga M. Olevsky2, Michael Fishbein3, Michael 4 3 3 1 Background: Long term survival results are not satisfactory following Selch , W Dean Wallace , Chi Lai , Anne Rorie 1 treatment for malignant pleural mesothelioma (MPM). However, a Thoracic Surgery, David Geffen School Of Medicine At UCLA, Los Angeles/ 2 small proportion of patients survive longer than 2 years. We analyzed CA/UNITED STATES OF AMERICA, Division Of Hematology-Oncology, the features of patients who survived longer than 2 years following David Geffen School Of Medicine At UCLA, Santa Monica/CA/UNITED 3 chemotherapy or multimodality treatment. STATES OF AMERICA, Pathology And Laboratory Medicine, David Geffen School Of Medicine At UCLA, Los Angeles/CA/UNITED STATES OF Methods: The patients with MPM who survived longer than 2 years AMERICA, 4Radiation Oncology, David Geffen School Of Medicine At UCLA, following chemotherapy or multimodality treatment (extrapleural Los Angeles/UNITED STATES OF AMERICA pneumonectomy, hemithoracic irradiation and chemotherapy) were identified from two prospective databases. Patient features including Background: The majority of patients with malignant pleural age, gender, side, histology, IMIG T and N stages (pretreatment for mesothelioma have a predominantly epithelioid histologic appearance. chemotherapy group and pathologic for multimodality treatment group), Tumors with sarcomatoid histology have an extremely poor prognosis. complete response/resection (microscopic) rate and survival were Due to encouraging experiences treating pleural sarcomas with induction compared using Student’s t-test and Kaplan Meier Survival analysis. chemotherapy followed by surgery and radiation, we sought to evaluate our results with a similar protocol for sarcomatoid mesothelioma.

Results: international mesothelioma interest group Methods: Following institutional IRB approval, we retrospectively reviewed our prospective thoracic database to identify patients with sarcomatoid- Chemotherapy Multimodality Treatment p predominant histology. All patients underwent pathological review by at (n=36) (n=12) least one of our panel of thoracic pathologists and were characterized No of pts/total 36/450 12/40 0.001 in terms of the percentage of sarcomatoid elements. Tumors with >50% sarcomatoid histology were classified as being sarcomatoid-predominant. Age 60 ± 11 51 ± 9 0.01 The treatment and outcomes of these patients were then investigated. Male/Female 15/21 4/8 NS Results: We identified 45 patients in our prospective database with Right/Left 21/15 5/7 NS sarcomatoid-predominant (>50% sarcomatoid elements) malignant pleural Histology 29/5/1 12 NS mesothelioma. Four (28.6%) patients were felt to have epithelioid histology (Epitelyal/ on original biopsy but ultimately were found to have sarcomatoid- MM/mixed) predominant biphasic tumors following resection. The mean age was 63.2 Complete 6/36 3/12 NS years, and 35 patients were male (77.8%) while 10 (22.2%) were female. response/ 26 (57.8%) were right-sided and 19 (42.2%) were left-sided tumors. Of the resection 45 patients, 2 are currently receiving induction chemotherapy and were excluded while 29 others (64.4%) were not felt to be surgical candidates, T1/2/3/4 8/10/13/5 2/7/2/1 NS developed or presented with metastatic disease, or opted to be treated Extrapleural 11/36 6/12 NS with palliative/supportive care. This resulted in a surgical cohort of 14 lymph node patients (31.1%) who underwent lung-sparing pleurectomy/decortication metastasis procedures. Compared to the overall group, these patients were slightly younger (mean age=58.2 years), more likely female (6 female=35.7%), Median (mo) 42/23 47,4 /28 NS and were evenly balanced right to left (7 right-sided=50% and 7 left- and 5-year sided=50%). Clinical T stage was T2 in 10=71.4%, T3 in 2=14.3%, T4 in Survival (%) 1=7.1%, and unclassified in 1. All were clinical N0,M0. Pathologic staging revealed T2 in 3=21.4%, T3 in 6=42.9%, and T4 in 4=28.6% while N2 The results are presented in the table. NS, not significant. disease was found in 4=28.6% and N0 in 10=71.4%. Two patients (14.3%) received no adjuvant therapy, and 4 (28.6%) only received an unknown Conclusion: The proportion of patients surviving longer than 2 years postoperative chemotherapy regimen, including 2 of the patients with appear to be larger in the multimodality treatment group. Although T original epithelioid biopsies. Of the remaining 8 patients, four recieved

iMig2012.org • Abstract Book 121 postoperative chemotherapy alone (two with interferon alpha followed by cisplatin/pemetrexed, one with cisplatin/gemcitibine, and one with ifosfamide/adrimycin). The last 4 patients received preoperative therapy with ifosfamide/adriamycin (3 patients) and cisplatin/pemetrexed/ Veglin (1 patient). Interestingly, 3/4 of these patients were noted to have pathological responses with 80-99% necrosis which is not often seen with standard cisplatin and pemetrexed chemotherapy. Although survival data is quite limited, survival roughly correlated with the existence of extensive necrosis following preoperative chemotherapy. Median survival of patients exhibiting >80% necrosis was 19.6 mos (with two patients still alive) compared to only 5.7 mos for the patient without extensive necrosis and 7.6 mos for the entire surgical group.

Conclusion: Sarcomatoid-predominant malignant pleural mesothelioma remains a difficult tumor to control. Optimal treatment strategies should employ multimodality approaches that stress systemic induction therapy with non-cisplatin/pemetrexed regimens. Salvage surgery may be an option for those patients who remain free of metastatic spread following poster sessions | september 13 induction therapy, particularly those with >80% necrosis. Results: The quarterly number of new patient visits and enrollment in Disclosure: No significant relationships. clinical trials since the formation of the Program 21 months ago is depicted in the figure. The total number of new patients seen, not all of whom were treated at our institution, was 182 and 60 (33%) of those patients were POSTER SESSION 3 September 13, 2012 11:30-12:30 treated on clinical trials. Conclusion: The percentage of patients being treated for MPM on P3.15: A DEDICATED MULTIDISCIPLINARY MESOTHELIOMA clinical trials in our institution is significantly higher than what is cited PROGRAM FOSTERS ENROLLMENT IN CLINICAL TRIALS as the typical clinical trial enrollment rate and is also significantly higher than the historical accrual to trials for MPM in our institution prior to Melissa J. Culligan1, Joseph Friedberg1, Adri Recio2, Mona Jacobs-Small3, formation of our dedicated multidisciplinary program. We attribute Karen Mudrick3, Steven M. Albelda4, Andrew R. Haas4, Keith Cengel5, this increased clinical trial enrollment rate to several features of the Stephen M. Hahn5, James Stevenson6, Charles B. Simone5, Corey Program: the increased influx of new patients, the mindset of the team Langer7, Evan Alley3, Daniel Sterman4 to support clinical trials and cross-pollination between disciplines, such 1Thoracic Surgery, University Of Pennsylvania, Philadelphia/PA/UNITED as the development of a new surgical pleural-access procedure to allow STATES OF AMERICA, 2University Of Pennsylvania Medical Center, enrollment of patients with fused chests (previously excluded) into a gene Philadelphia/PA/UNITED STATES OF AMERICA, 3Medicine, University therapy trial. We conclude that one of the benefits of a dedicated MPM Of Pennsylvania/UNITED STATES OF AMERICA, 4Division Of Pulmonary, program is that it fosters clinical trial enrollment. Allergy And Immunology, University Of Pennsylvania School Of Medicine, 5 Philadelphia/UNITED STATES OF AMERICA, Radiation Oncology, University Disclosure: No significant relationships. Of Pennsylvania/UNITED STATES OF AMERICA, 6Taussig Cancer Institute, Cleveland Clinic Foundation/UNITED STATES OF AMERICA, 7Medicine, University Of Pennsylvania, Philadelphia/UNITED STATES OF AMERICA POSTER SESSION 3 September 13, 2012 11:30-12:30 Background: Clinical trials are the mechanism for both introducing international mesothelioma interest group innovative therapies and establishing the standard-of-care in cancer P3.16: P/D + HITHOC (HYPERTHERMIC INTRATHORACIC treatments, yet the number of patients treated on clinical trials is typically CHEMOPERFUSION) IS COMPARABLE TO EPP IN THE cited as less than 5%. Malignant pleural mesothelioma (MPM), a cancer for which novel therapies are desperately needed and for which the standard- MULTIMODALITY TREATMENT OF PATIENTS WITH MALIGNANT of-care is limited to palliative chemotherapy, is clearly a cancer for which PLEURAL MESOTHELIOMA (MPM) clinical trials should be encouraged. In an effort to optimize the care of MPM patients at our institution, we formed a multidisciplinary program Uwe Gruetzner, Michael Lindner, Martin E. Eichhorn, Juergen Sklarek, dedicated solely to treatment of patients with pleural malignancies. From Sandra Feske, Rudolf A. Hatz the outset, one of the goals of the Program was to maximize enrollment in Department Of Thoracic Surgery, Munich Center For Thoracic Surgery, clinical trials. This report details the results of this effort. Asklepios Kliniken Muenchen-Gauting, Gauting/GERMANY

Methods: In September 2010 the Penn Mesothelioma and Pleural Program Background: The objective was to evaluate the overall survival rates was formed. The format of the Program is to meet weekly to review in patients with resectable MPM treated by EPP or P/D + HITHOC in a patient cases, consider all potential treatment options amongst the team retrospective non-randomized consecutive study. and then have each discipline meet with the patients to discuss potential treatment options. Typically each patient is seen by: pulmonary medicine/ Methods: From 10/2002 until 06/2011, a total of 56 patients were immunotherapy, thoracic surgery, medical oncology, radiation oncology/ consecutively treated by Extrapleural Pneumonectomy (EPP) or open photodynamic therapy. Although patients are treated off protocol, every Pleurectomy/Decortication (P/D) in combination with HITHOC using attempt is made to treat patients on a clinical trial, which include: surgery- cisplatin (40mg/l) and doxorubicin (20 mg/l) in a total volume of 5000 ml based multimodal therapy, chemotherapy, immunotherapy. normal saline for 90 minutes at 42°C using a Performer HT (RanD, Medolla, Italy). Starting in 11/2009 P/D+HITHOC was first offered as treatment option for patients not qualifying for EPP because of age and/or cardio- respiratory conditions.

Results: There were 28 patients in each group. EPP, P/D and all chemoperfusions were completed as planned. The P/D group consists of 22 male and 6 female patients aged 53-80 (median 71) years. One patient died from pulmonary embolism. We observed cardiac arrhythmias in 8/28, respiratory failures in 6/28 and reoperations in 3/28 patients (1 thoracic duct lesion, 1 empyema, 1 persistent air leak). Mean days in ICU

iMig2012.org • Abstract Book 122 were 3, in hospital 26 days respectively. No potential side effects of the Disclosure: No significant relationships. chemoperfusion were observed, such as wound healing disorders, nausea, or significant rise in creatinine or drop in WBC. EPP group consisted of 24 male and 4 female patients aged 30-71 (median 57) years. 15/28 pts. suffered from cardial and 2/28 from pulmonary complications. POSTER SESSION 3 September 13, 2012 11:30-12:30 5 reoperations were performed (3 empyema, 1 gastric herniation, 1 bronchus leakage). Mean number of days in ICU were 5, and 34 in hospital P3.18: PLEURECTOMY/DECORTICATION, HYPERTHERMIC respectively. The pericardium and diaphragm were resected and replaced PLEURAL LAVAGE WITH POVIDONE-IODINE FOLLOWED BY all EPP and in 3/28 and 3/28 P/D-patients respectively. Median survival ADJUVANT CHEMOTHERAPY: SURVIVAL ANALYSIS IN 65 in P/D group was 18,6 months (95% CI 17,6-19,6 months) and 22,8 months CONSECUTIVE PATIENTS (95% CI 12,8-32,3 months) in EPP group. P-value (Log Rank) was 0,369 (n.s.). Andrea Bille’1, Lawrence Okiror1, James Spicer2, Jeremy P. Steele2, David Landau2, Henry Taylor2, Loic Lang-Lazdunski1 Conclusion: The oncological results and overall survival rate for patients 1Department Of Thoracic Surgery, Guy’s Hospital, London/UNITED treated with P/D + HITHOC (Cis-Doxo) is comparable to EPP. It represents KINGDOM, 2Department Of Oncology And Hematology, Guy’s Hospital, a reasonable treatment option for patients not eligible to undergo EPP London/UNITED KINGDOM for age and/or cardiopulmonary reasons. Taking into account the shorter stay in hospital, the lesser rate of reoperations and the sparing of lung Background: Radical surgery remains controversial in malignant pleural poster sessions | september 13 parenchyma only marginally affected by the tumor, it is warranted if P/D mesothelioma (MPM). Our goal was to examine the results of pleurectomy/ + HITHOC is also an option in patients where today EPP is still standard of decortication (P/D) with hyperthermic pleural lavage with povidone- care. iodine followed by prophylactic radiotherapy radiotherapy and adjuvant chemotherapy in patients with malignant pleural mesothelioma. Disclosure: No significant relationships. Methods: Prospective study of patients undergoing P/D and hyperthermic pleural lavage with povidone-iodine, prophylactic radiotherapy (21 Gy in 3 fractions) and adjuvant chemotherapy at our institution. All procedures POSTER SESSION 3 September 13, 2012 11:30-12:30 were performed by the same surgeon and patients were treated by the same team. P3.17: HYPERTHERMIC INTRATHORACIC CHEMOPERFUSION (HITHOC) IN COMBINATION WITH PLEURECTOMY / Results: Between October 2004 and March 2012, sixty five consecutive DECORTICATION (P/D) FOR TREATMENT OF MALIGNANT PLEURAL patients were offered this multi-modality therapy. Fifty four patients were MESOTHELIOMA (MPM) male, and 11 female. The median age at operation was 63 year (range 45-74). The 30-day mortality was nil. Twelve patients (18.5%) experienced Uwe Gruetzner1, Michael Lindner1, Martin E. Eichhorn1, Sandra Feske1, postoperative complications: 6 patients (9.2 %) had a persistent air Juergen Sklarek1, Thomas Duell2, Rudolf A. Hatz1 leak, 4 patients (6.1 %) had chylothorax requiring surgical intervention, 1Department Of Thoracic Surgery, Munich Center For Thoracic Surgery, one patient had ARDS (1.5 %), one patient (1.5%) had pneumonia and Asklepios Kliniken Muenchen-Gauting, Gauting/GERMANY, 2Department one patient (1.5%) had postoperative empyema. All patients received For Pneumological Oncology, Asklepios Kliniken Muenchen-Gauting, prophylactic radiotherapy and adjuvant chemotherapy. Three patients Gauting/GERMANY received 2 cycles, 62 patients received 4-6 cycles of cisplatin-based chemotherapy. Second line chemotherapy (vinorelbine or pemetrexed) or Background: Our objective was to evaluate the oncological results of P/D + phase 1-2 trials were routinely offered to patients with performance status HITHOC (Cis-Doxo) in elderly patients with potentially resectable MPM. 0-2 progressing after first line chemotherapy. At last follow-up, thirty six of 65 patients were alive (median follow-up 21 months, range 4.4-76.4 Methods: From 11/2009 until 06/2012, a total of 46 patients were treated months). Twenty eight patients were alive with no evidence of disease international mesothelioma interest group by open pleurectomy and complete decortication of the lung to remove recurrence, 8 were alive with disease recurrence and 29 patients had died all resectable tumor mass. After closing the chest cavity a Performer HT of disease progression. The overall 1-year survival was 90%, 2-year survival (RanD, Medolla, Italy) was used for chemoperfusion with cisplatin (40mg/l) was 62.5 % and 5-year survival 33.4%. At univariate analysis, complete and doxorubicin (20 mg/l) in a total volume of 5000 ml normal saline for macroscopic resection (p=0.002) and epithelioid histology (p=0.009) were 90 minutes at 42°C. associated with longer survival. Patients undergoing complete macroscopic resection (R0-R1) had 1-year, 2-year and 5-year survival of 100%, 84.8% Results: 46 patients (37 male / 9 female), age 52-81 (median 70) and 43.2%, respectively. Patients who underwent incomplete macroscopic years were successfully treated. Surgery and all chemoperfusions were resection (R2) had 1-year, 2-year and 5-year survival of 76%, 34.8% and completed as planned. The pericardium and diaphragm were resected and 21.8%, respectively. Patients with epithelioid subtype had a 1-year, 2-year replaced in 7/46 and 4/46 pts respectively. Time under anesthesia was and 5-year survival of 100%, 74.8% and 44.7%, respectively. Patients with between 6:05 and 10:10 (median 7:10) hrs, extubation was possible right biphasic/sarcomatoid subtype had 1-year, 2-year and 5-year survival of after the operation in all but one case. Postoperative treatment in the ICU 68.4%, 34.2% and 13.7%, respectively. Patients with epithelioid subtype was necessary for 1-11 (median 1) days. One patient died from pulmonary and complete macroscopic resection (n=31) had 1-year, 2-year and 5-year embolism. No serious side effects of the chemoperfusion were observed, survival of 100%, 86.2% and 46%, respectively. such as wound healing disorders, nausea, or significant rise in creatinine or drop in WBC. We observed cardiac arrhythmia n=11/46 (24%), respiratory Conclusion: Pleurectomy/decortication with hyperthermic pleural failure n=10/46 (22%), pneumonia n=7/46 (15%) and secondary air leaks lavage with povidone-iodine, prophylactic radiotherapy and adjuvant n=10 (22%). 4 reoperations occurred (1 thoracic duct lesion, 1 empyema, 2 chemotherapy is a safe and well tolerated multi-modality therapy. Patients persistent air leaks) Follow-up time was 1 to 31 (mean 9,1) months. 38/46 with epithelioidsubtype in whom complete macroscopic resection can be (83%) patients are still alive. Mean overall survival time was 8,2 months. achieved experience the best outcomes.

Conclusion: P/D + HITHOC (Cis-Doxo) proofed to be a successful surgical Disclosure: No significant relationships. treatment option for MPM. It can be offered to patients not eligible to undergo EPP for age and/or cardiopulmonary reasons. Despite the long time under anesthesia and high dose local chemotherapy patients are doing remarkably well after surgery. The oncological results are comparable to EPP. It should be warranted to investigate if P/D + HITHOC is also an option in patients where today EPP is still considered the gold standard of care.

iMig2012.org • Abstract Book 123 POSTER SESSION 3 September 13, 2012 11:30-12:30 procedure in which intraoperative cisplatin chemotherapy is commonly administered. Because of the high risk of AKI and the devastating P3.19: PHARMACOKINETIC STUDY OF HYPERTHERMIC consequences of chronic kidney disease and end stage renal disease in INTRATHORACIC CHEMOTHERAPY (HITHOC) FOLLOWING this cancer cohort, we studied the ability of early postoperative changes in PLEURECTOMY AND DECORTICATION serum creatinine (sCr) to predict sustained kidney injury. We hypothesized that acute sCr elevation, within 24-48 hours after surgery, can predict Michael Lindner1, Ina Koch1, Uwe Gruetzner1, Martin Eichhorn1, Thomas sustained, clinically significant decline in kidney function defined as 50% Buech2, Rudolf Hatz1 increase in sCr from baseline, 2-4 weeks after surgery. 1Thoracic Surgery Center, Comprehensive Pneumology Center Ludwig Maximilians University Munich Asklepios Clinics Munich - Gauting, Munich Methods: A prospective, observational cohort of patients undergoing - Gauting/GERMANY, 2Walther-Straub-Institute For Pharmacology And surgical resection (extrapleural pneumonectomy) for the treatment of Toxicology, Ludwig-Maximilians-University Munich, Munich/GERMANY malignant pleural mesothelioma at the Brigham and Women’s Hospital between 1998 and 2009 were studied. Receiver operator characteristic Background: Cytoreductive surgery in combination with hyperthermic (ROC) curves were generated to examine the diagnostic ability of 24- pleural perfusion with chemotherapy agents is an ideal multimodality and 48-hour changes in sCr over baseline to identify sustained kidney approach to treating malignant pleura mesothelioma. The aim of this study injury in the derivation cohort (n=279) and tested in the validation cohort is to investigate the pharmacokinetic behavior of cisplatin and doxorubicin (n=207). The performance of our derived criteria was compared to various during this procedure. other criteria used to characterize AKI to predict the risk of sustained poster sessions | september 13 kidney injury using the net reclassification index (NRI) and integrated Methods: After pleurectomy and decortication via anterolateral discrimination improvement (IDI). thoracotomy a 42° Celsius hyperthermic solution of a 5000ml saline containing 200mg Cisplatin and 100mg Doxyrubicin was perfused into Results: Sustained kidney injury occurred in 8.9% (n=25) of patients in the thoracic cavity for 90 minutes. Simultaneous samples to analyze the the derivation and 10.1% (n=21) in the validation cohort. A 59% or greater concentration of chemotherapy agents were drawn from the hyperthermic increase in sCr (1.59 fold) at 48 hours after surgery was most predictive of pump at 0, 10, 30, 60, 90 minutes after cisplatin and doxorubicin had been sustained kidney injury (AUCROC= 0.798; sensitivity of 68% and specificity perfused. 3 milliliter blood samples were subsequently obtained at minute of 87%). When compared to various other AKI criteria, we found that our 0, 10, 30, 60, 90 and day 1, 2 and 4. Doxorubicin fluid and serum levels prediction model had the highest c-statistic; and when compared to the were analyzed by high performance liquid chromatography. Cisplatin was RIFLE criteria the difference was statistically significant (p<0.001). Among measured by a flameless atomic absorption spectrometry. Clinical datasets other AKI definitions, we found that sCr increase of 0.3 mg/dl in 24 hours including perioperative morbidity were recorded. or 0.5 mg/dl increase in 48 hours (Waikar and Bonventre criteria) also reliably predict sustained kidney injury. Results: Recently, 46 hyperthermic intrathoracic chemotherapy (HITHOC) procedures have been performed and from 9 patients a complete dataset Conclusion: In this study we identified a cohort, which developed was analyzed. The intrapleural administration of cisplatin and doxorubicin sustained kidney injury (defined as doubling of sCr present at 2-4 weeks consisted in an increased exposure to total platinum and doxorubinol of after surgery). We found that development of clinically significant sustained the pleural cavity without increasing the systemic circulation. Local toxicity kidney injury can be predicted by acute postoperative sCr elevation in was not seen after the procedure. The AUC (0-90 minutes) and their mean patients treated for mesothelioma where a 59% (1.59 fold) sCr elevation ratio AUC (regional/systemic values) show that the absorption was greater at 48 hours was the best predictor of sustained kidney injury (positive after neoadjuvant therapy. The mean resorption intake was 8.5% (4.8-16.1) predictive value of 41%; negative predictive value of 96%). for doxorubicin and 18.6 % (11.5-22.1) for Cisplatin.The serum Cmax was median 1.73 μg/ml for cisplatin and 90.9 ηg/ml for doxorubicin. Disclosure: No significant relationships.

Conclusion: After an aggressive cytoreductive lung sparing surgery hyperthermic chemotherapy is feasible, save and pharmacologically international mesothelioma interest group favorable.This study should be a step forward to delivering a standardized POSTER SESSION 3 September 13, 2012 11:30-12:30 HITHOC protocol. P3.21: TWO SIMULTANEOUS, CONSECUTIVE, NON-RANDOMIZED Disclosure: No significant relationships. COHORTS OF OPERABLE PATIENTS WITH MALIGNANT PLEURAL MESOTHELIOMA (MPM) RECEIVING INDUCTION CHEMOTHERAPY FOLLOWED BY EITHER EXTRAPLEURAL PNEUMONECTOMY (EPP) AND POSTOPERATIVE RADIOTHERAPY POSTER SESSION 3 September 13, 2012 11:30-12:30 OR PLEURECTOMY/DECORTICATION ALONE, RESPECTIVELY. THE P3.20: POSTOPERATIVE CREATININE INCREASE PREDICTS DANISH EXPERIENCE. SUSTAINED KIDNEY INJURY IN PATIENTS WITH MALIGNANT Jens Benn Sorensen1, Jesper Ravn2, Claus Kristensen1, Anne K. MESOTHELIOMA UNDERGOING SURGICAL RESECTION Berthelsen3, Annika Loft4 1Dept. Oncology, Finsen Centre/National University Hospital, Copenhagen/ 1 1 2 K Annette Mizuguchi , Aya Mitani , Sushrut Waikar , David J. DENMARK, 2Department Of Thoracic Surgery, 2151, Rigshospitalet, 3 2 4 Sugarbaker , Joseph Bonventre , Gyorgy Frendl Copenhagen University Hospital/DENMARK, 3Radiation Oncology, National 1 Anesthesiology Perioperative And Pain Medicine, Brigham And Women’s University Hospital, /DENMARK, 4Clinical Physiology, National University 2 Hospital, Boston/MA/UNITED STATES OF AMERICA, Medicine, Renal Hospital/DENMARK Division, Brigham And Women’s Hospital, Boston/MA/UNITED STATES OF 3 AMERICA, Division Of Thoracic Surgery, Brigham And Women’s Hospital, Background: Trimodality treatment combining chemotherapy, extrapleural 4 Boston/MA/UNITED STATES OF AMERICA, Anesthesiology, Perioperative, pneumonectomy (EPP) and postoperative radiotherapy may result in cure And Pain Medicine, Brigham And Women’s Hospital, Boston/MA/UNITED (5-year survival) in some MPM patients but prognosis is generally poor STATES OF AMERICA and treatment is affiliated with morbidity and risk of treatment related deaths. Pleurectomy/decorticatio (P/D) is a less comprehensive operative Background: Acute kidney injury (AKI) leads to increased morbidity procedure sparing the lung and with less morbidity but theoretically with and mortality and progression to chronic kidney injury via a subchronic, dubious curative potential. Results from two such simultaneous treatment mid-term kidney injury (sustained kidney injury) phase is a frequent cohorts in Denmark are given (not randomized). consequence of AKI. The incidence of severe AKI (serum creatinine elevation > 3 times upper limit of normal) is nearly 10% in patients Methods: Induction chemotherapy for both groups were 3-6 courses undergoing surgical resection of malignant pleural mesothelioma, a of platinum-based doublet chemotherapy. F18-FDG-PET/-CT scan fused

iMig2012.org • Abstract Book 124 imaging restaging and preoperative mediastinoscopy was subsequently not progress on CT scan will then be randomised to: (1) up to 4 further performed at Rigshospitalet, Copenhagen, Denmark where also all surgery cycles of Pemetrexed-Platinum chemotherapy; or (2) ePD followed by up was performed. All patients had T1-3N0-1M0, age ≤70 years, performance to 4 further cycles of post-operative Pemetrexed-Platinum chemotherapy. status 0-1, and no major comorbidities. EPP was performed in patients Major inclusion criteria are: histological confirmation of MPM, and tumour having epitheloid subtype and lung function test allowing pneumonectomy. confined to one hemithorax. Major exclusion criteria are: refusal to give In that case EPP was followed by postoperative radiotherapy 56 Gy in informed consent; refusal to accept randomisation; disease extent deemed 30 fractions, 5F/W, and radiotherapy constraints for contralateral lung non-resectable; inadequate pulmonary, cardiac, hepatic or renal reserve. was V20<15%, V10<50%, and MLD<12 Gy (trimodal treatment). P/D was performed in patients having biphasic or sarcomatoid subtypes, poor lung Results: This is the first time the final trial plan for MARS-2 has been function test, or cardial comorbidity not allowing EPP but allowing lesser presented at an international meeting. The trial is now under submission surgery. with Cancer Research UK (CRUK) for resource approval. The full trial protocol will be discussed in detail. Results: In the EPP group of 28 Danish patients included 2006-2011 there were 2 cases of fatal radiation pneumonitis (7%) and no 30- Conclusion: There is a pressing need for better data on the benefits of days postoperative deaths. Median postoperative hospital stay was 15 surgery in MPM. Audits of thoracic surgical centres in the UK show that days (range 8-29 days). The P/D group of 34 patients likewise had no over 100 MPM patients undergo a pleurectomy every year. This trial aims postoperative fatalities and median postoperative hospital stay was 7 days to demonstrate whether this type of operation can offer survival and/or (range 3-27 days). Median survival rates for EPP and P/D groups were 31.9 quality-of-life benefits to patients with MPM. The trial plan is in submission poster sessions | september 13 and 28.4 months, respectively. 5-years survival rates were 16% and 27%, for resources. If funded and successful MARS-2 will be the largest and most respectively. detailed surgical trial in MPM.

Conclusion: EPP is a safe procedure when performed in high-volume Disclosure: No significant relationships. institution but affiliated with longer postoperative hospital stay than P/D. Especially postoperative radiotherapy remains a high-risk part of the trimodality treatment which should be further explored. Cure seems possible in about 1/6 of the patients. P/D is a safe procedure without treatment related deaths and the 5-year survival in about ¼ of patients may suggest that cure may be possible with this treatment when preceded by induction chemotherapy. The relative benefit of these two treatment options (trimodal treatment with EPP, or P/D) cannot be settled from these results which were offered somewhat different patient populations. However, a randomized trial might be considered.

Disclosure: No significant relationships.

POSTER SESSION 3 September 13, 2012 11:30-12:30

P3.22: A RANDOMISED TRIAL OF EXTENDED PLEURECTOMY AND DECORTICATION (‘EPD’) WITH PEMETREXED AND PLATINUM CHEMOTHERAPY VERSUS PEMETREXED AND PLATINUM ALONE IN PATIENTS WITH MALIGNANT PLEURAL MESOTHELIOMA international mesothelioma interest group (‘MARS-2’).

Jeremy P. Steele1, David A. Waller2, John Edwards3, Tom Treasure4, Sanjay Popat5, Riyaz Shah6, Robert C. Rintoul7, Julian Peto8, Eric Lim9 1Medical Oncology, St Bartholomew’s Hospital, London/UNITED KINGDOM, 2Glenfield Hospital/UNITED KINGDOM, 3University Hospitals Sheffield/UNITED KINGDOM, 4Ucl/UNITED KINGDOM, 5Royal Marsden Hospital/UNITED KINGDOM, 6Kent Cancer Centre/UNITED KINGDOM, 7Papworth Hospital/UNITED KINGDOM, 8University Of London/ UNITED KINGDOM, 9Royal Brompton Hospital/UNITED KINGDOM

Background: The role of radical surgery in malignant pleural mesothelioma (MPM) is not clearly defined. There has been only one randomised trial of surgery (‘MARS-1’), which showed no benefit for extra- pleuralpneumonectomy (EPP) compared to the non-surgical control arm. MARS-1 recruited more slowly than anticipated and did not progress to the larger scale trial originally planned. Extended pleurectomy and decortication (ePD) is a less extensive operation that does not mandate pneumonectomy, with probable lower morbidity and mortality than EPP. More patients with MPM may be suitable for ePD than were for EPP, enabling more rapid recruitment and randomisation.

Methods: The initial phase of the trial with be a randomised feasibility study. The primary endpoint is the ability to randomise 50 patients in 24 months. Secondary endpoints are: survival from time of randomisation and quality-of-life (assessed by the QLQ 30 and LC-13 modules). If fewer than 30% of suitable patients accept randomisation, it is unlikely that the larger study would start. The treatment plan is as follows: patients with MPM will be registered on MARS-2 and will receive 2 cycles of Pemetrexed-Platinum chemotherapy [cisplatin or carboplatin are acceptable]; patients who do

iMig2012.org • Abstract Book 125 Poster Session 3 Immunology September 13, 2012 11:30-12:30

POSTER SESSION 3 September 13, 2012 11:30-12:30 can be used as to target residual disease, but these too are unlikely to be curative. There is renewed interest in the use of immunotherapy to P3.23: IMPACT OF THREE DIFFERENT SURGICAL PROCEDURES treat mesothelioma as new modalities have been developed and immune FOR MALIGNANT PLEURAL MESOTHELIOMA ON IMMUNOLOGIC targeting therapies have been found to be useful in other cancers. Recent RESPONSES work form our laboratory and others, has demonstrated that specific

immunotherapies can engage the immune system with tumour. These poster sessions | september 13 Vincenzo Ambrogi, Federico Tacconi, Francesco Sellitri, Tommaso C. therapies are typically more effective when used in conjunction with Mineo standard treatment protocols such as surgery or chemotherapy. Here we Thoracic Surgery Dept., Tor Vergata University, Rome/ITALY describe the development of a prime boost (P/B) anti-tumour vaccination protocol that when combined with debulking surgery and removal Background: Intentional surgical radical procedures for malignant pleural of regulatory T cells improved survival outcome in a mouse model of mesothelioma (MPM) often achieve disappointing results. Furthermore mesothelioma. surgical stress may have detrimental effects on immune response. We comparatively evaluated changes of immunologic response by assessing Methods: Using our established mouse model of mesothelioma, AB1-HA lymphocyte subsets and cytokines in patients with MPM after three tumour bearing mice received influenza A PR8 (prime) and rMVAHA (boost) different operations. anti-tumour vaccinations, either before (neoadjuvant) or after (adjuvant) 75% debulking surgery. Tumour growth was monitored and immunological Methods: Between October 2004 and May 2012, 30 patients with parameters assessed by multicolour FACS. epithelial MPM underwent biopsy-pleurodesis in videoassisted thoracoscopy (n=15), open pleurectomy-decortication (n=8) and Results: Neoadjuvant P/B vaccination alone or in combination with 75% extrapleural pneumonectomy (n=7). Total lymphocyte count and debulking surgery induced a significant increase in splenic tumourspecific percentage changes of lymphocyte subsets including natural-killer cell and CD8 T cells as well as significant increases in the proportion, activation and cytokines (i.e interleukin-6 and -10) were evaluated by two-way analysis proliferation status of peripheral CD8 T cells relative to other treatment of variance test for repeated measures pre and postoperative days 1, 7 groups. However, a significant delay in tumour growth was only observed and 14. The Mann-Whitney test was performed at each time point only for when neoadjuvant P/B vaccination was combined with debulking surgery; significant parameters at between-group analysis of variance. although this was not sufficient to control tumour outgrowth and cure the animals. Targeted depletion of CD8 and CD4 T cells demonstrated that Results: Preoperative data were relatively homogeneous in all groups. CD8 T cells were essential for the delay in tumour growth caused by the Both the pleurectomy-decortication and extrapleural pneumonectomy combination treatment. Interestingly, depletion of CD4 T cells during P/B groups disclosed a significantly lower median (interquartile range) vaccination enhanced the survival outcome of surgery + P/B vaccination proportion of natural-killer cells as compared with the biopsy-pleurodesis with 60% of these mice remaining tumour free for > 60 days post-surgery. group on postoperative day 1 [5%(3-8%) and 4%(3-5%) Vs 12%(8-14%), Immunological memory was confirmed as tumour specific CD8 T cells were international mesothelioma interest group p<0.003) and day 7 [7%(4-10%) and 3%(2-5%) Vs 11%(8-21%), p<0.02]. detected in the LN and spleens of surviving mice >60 days post tumour Total lymphocyte count significantly decreased only in the extrapleural rechallenge. pneumonectomy (p<0.00001). No difference was found in the remaining lymphocyte subsets. Interleukin-6 and -10 were significantly increased Conclusion: Anti-tumour P/B vaccination induced tumourspecific after pleurectomy-decortication (postoperative day 7: p<0.03 and immunity resulting in delayed tumour growth when combined with p<0.008; day 14: p<0.05 and p<0.02) and extrapleural pneumonectomy debulking surgery. Depletion of CD4 T cells during neoadjuvant P/B (day 7:p<0.01 and p<0.003, day 14:p<0.04 and p<0.01). vaccination enhanced the therapeutic efficacy leading to cures in 60% of treated mice, suggesting that vaccine induced antitumour immunity Conclusion: Biopsy-pleurodesis resulted in a lesser impact on is restrained, probably by CD4+ regulatory T cells. Based on these findings postoperative immunologic response than more aggressive procedures and we are investigating whether combining novel immunotherapies with this should be considered in choosing therapeutic strategy. conventional treatments in the absence of immunological restrainers may generate effective therapy for mesothelioma. Disclosure: No significant relationships. Disclosure: No significant relationships.

POSTER SESSION 3 September 13, 2012 11:30-12:30 POSTER SESSION 3 September 13, 2012 11:30-12:30 P3.24: COMBINING PRIME-BOOST ANTI-TUMOUR VACCINATION WITH DEBULKING SURGERY FOR THE TREATMENT OF P3.25: MEASLES VIRUS VACCINE-INFECTED TUMOR CELLS MALIGNANT MESOTHELIOMA INDUCE PLASMACYTOID DENDRITIC CELL MATURATION AND TUMOR ANTIGEN CROSS-PRESENTATION: POTENTIAL Scott Fisher, Amanda Cleaver, Andrea Khong, Theresa Connor, Ben APPLICATION TO MESOTHELIOMA TREATMENT. Wylie, Daphne Lakhiani, Bruce W.S. Robinson, Richard Lake University Of Western Australia School Of Medicine And Pharmacology, Marc Gregoire1, Jean-baptiste Guillerme1, Nicolas Boisgerault1, David National Centre For Asbestos Related Diseases, Perth/WA/AUSTRALIA Roulois1, Frédéric Tangy2, Jean-françois Fonteneau1 1Inserm-892/Cnrs-6299, Institute Of Therapeutic Research, Nantes/ Background: Malignant mesothelioma is a highly aggressive cancer with FRANCE, 2Laboratoire De Génomique Virale Et Vaccination, Pasteur a very poor prognosis. Debulking surgery is often used as the principal Institute, Paris/FRANCE therapy but is seldom curative. Adjuvant chemotherapy or radiotherapy

iMig2012.org • Abstract Book 126 Background: Plasmacytoid dendritic cells (pDC) are antigen-presenting Methods: Pleural fluid of 140 mesothelioma patients was collected. cells specialized in antiviral response. The measles virus vaccine (MV) For each sample the total cellular amount was determined. Cytospins was recently proposed as an antitumor agent to target and specifically were prepared and cell pellets were stored for later analysis. kill tumor cells without infecting healthy cells. Here, we investigated in Immunohistochemical staining for tumor cells, stromal cells and immune vitro the effects of MV-infected tumor cells on phenotype and functions of cells was performed on the cytospins. Cytokine profiles will be determined. human pDC. Clinical data and survival data are currently being collected and will be compared to the immunological data. Methods: Mesothelioma Meso13 cell line, melanoma M18 and pulmonary adenocarcinoma A549 cell line were cultured in the presence of Live- Results: Preliminary data show distinct differences in immune cell attenuated Schwarz strain Measles Virus (MV vaccine) or irradiated by UV-B composition of pleural fluid of mesothelioma patients. The clinical to induce cell death (apoptosis). Monocytes (Mo-DC) and plasmacytoid DC significance of these data will be determined in the months to come. (pDC) were purified from Peripheral Blood Mononuclear Cells (PBMCs) of healthy donors by counter-flow centrifugal elutriation at a purity of 95%. Conclusion: We are currently undertaking a study to investigate the Mo-DC, were generated in vitro after 6 days of culture in RPMI containing cellular composition of pleural fluid and its potential prognostic value in 2% human albumin, 1000 U/ml rhGM-CSF and 200 U/ml rhIL-4. pDC were patients with mesothelioma. maintained in culture with 20ng/ml of rhIL-3 and activated in vitro with a TLR-7 agonist R848 (5µg/ml). Immature Mo-DC or pDC were cultured in the Disclosure: No significant relationships. presence of MV-infected or UV-irradiated tumor cells (1 DC : 1 tumor cell). poster sessions | september 13 Phenotype of pDC and Mo-DC was determined by immunofluorescence followed by flow cytometry. After 18h, Mo- or pDC were co-cultured with the HLA-A*0201/NYESO-1(156-165) specific CD8+ T-cell clone, M117.167, for 6h in presence of Brefeldin-A. As a positive control, we used Mo-DC or pDC pulsed 1h with 0.1 or 1µM of NYESO-1 (156-165) peptides and washed. IFN-γ production by the T cell clone was analyzed by flow cytometry with a gate on CD8+ T cells using mAb specific for IFN-γ.

Results: We studied maturation, cytokine production and tumor-antigen cross-presentation by pDC, exposed either to the virus alone, or MV- infected or UV-irradiated tumor cells. We found that MV-infected cells induced pDC maturation with a strong production of IFN-α, whereas UV-irradiated tumor cells were unable to activate pDC. This production of IFN-a was triggered by the interaction of MV ssRNA with TLR7. We also observed that MV-infected tumor cells were phagocytosed by pDC. Interestingly, we observed cross-presentation of the tumor antigen NYESO-1 to a specific CD8+ T-cell clone, when pDC were cocultured with MV-infected tumor cells, whereas pDC were unable to cross-present NYESO-1 after co-culture with UV-irradiated tumor cells.

Conclusion: Altogether, our results suggest that the use of MV in antitumor virotherapy induces immunogenic tumor cell death, allowing pDC to spontaneously mature, to produce high amount of IFN-a, and to cross- present tumor antigen representing therefore a way to recruit cytotoxic T cells in the anti-tumor immune response. Thus the mealses virus vaccine could be used as an immune strategy to treat mesothelioma patients, international mesothelioma interest group inducing an activation of specific cytotoxic T cell populations afer activation by the antigen presenting cells: monocyte and plasmacytoid dendritic cells.

Disclosure: No significant relationships.

POSTER SESSION 3 September 13, 2012 11:30-12:30

P3.26: IMMUNOLOGICAL CHARACTERISTICS OF PLEURAL FLUID IN MALIGNANT MESOTHELIOMA

Lysanne A. Lievense, Joost P.J.J. Hegmans, M E.H. Lambers, H C Hoogsteden, J G J V Aerts Pulmonary Medicine, Erasmus Medical Centre, Rotterdam/NETHERLANDS

Background: Dyspnoe is a common characteristic of patients with malignant pleural mesothelioma and thoracentesis is indicated to relieve symptoms. The procedure is relatively easy and the obtained pleural fluid might reflect the tumor micro-environment. The effusion consists of varying amounts of tumor cells, stromal cells, and numerous types of immune cells. These cellular populations and protein content of the effusion could correlate with the aggressiveness of a tumor. Immune cells can possess either immune stimulatory capacity (eg CD8-cells and NK- cells) or immune suppressive capacity (eg tumor-associated macrophages or regulatory T-cells). The balance between these stimuli may determine tumor progression. Therefore, the proteomic and immunological characteristics of pleural fluid are subject of this study.

iMig2012.org • Abstract Book 127 Poster Session 3 Radiation Therapy and Biology September 13, 2012 11:30-12:30

POSTER SESSION 3 September 13, 2012 11:30-12:30 POSTER SESSION 3 September 13, 2012 11:30-12:30

P3.27: INTERNATIONAL SURVEY OF RADIATION TOXICITY P3.28: PRESCRIPTION DOSE PREDICTION MODEL FOR FOLLOWING HEMITHORACIC INTENSITY-MODULATE HEMITHORACIC INTENSITY-MODULATED RADIATION THERAPY RADIOTHERAPY IN PATIENTS WITH MALIGNANT PLEURAL IN MESOTHELIOMA PATIENTS WITH TWO INTACT LUNGS MESOTHELIOMA poster sessions | september 13 Li C. Kuo1, Ellen Yorke1, Amanda Mclane2, James Mechalakos1, Abraham Malcolm Feigen1, Christopher Kelsey2, Vivek Mehta3, John Cho4, J. Wu2, Kenneth E. Rosenzweig3, Andreas Rimner2 Philippe Giraud5, Marc A. Mahe6, Marta Scorsetti7, Sandro Tonoli8, 1Department Of Medical Physics, Memorial Sloan-Kettering Cancer Center, Florian Sterzing9, Jerome Krayenbuhl10, Aaron M. Allen11 New York/NY/UNITED STATES OF AMERICA, 2Department Of Radiation 1Radiation Oncology Center, Austin Health, Heidelberg West/VIC/ Oncology, Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED AUSTRALIA, 2Dept Of Radiation Oncology, Duke University Medical STATES OF AMERICA, 3Department Of Radiation Oncology, Mount Sinai Center, NC/UNITED STATES OF AMERICA, 3Dept Of Radiation Oncology, Medical Center, New York/NY/UNITED STATES OF AMERICA Swedish Cancer Institute, Seattle/UNITED STATES OF AMERICA, 4Toronto General Hospital And Princess Margaret Hospital, Toronto/ON/ Background: Intensity-modulated radiation therapy (IMRT) to the CANADA, 5Radiation Oncology, Hopital Europeen Georges Pompidou, entire hemithoracic pleura is a promising new strategy for patients with Paris/FRANCE, 6Radiation Oncology, Centre René Gauducheau, Nantes/ unresectable malignant pleural mesothelioma (MPM) or after pleurectomy/ FRANCE, 7Dept Of Radiation Oncology, Instituto Clinico Humanitas, Milan- decortication (P/D). The radiation tolerance of the normal lungs is the main rozzano/ITALY, 8Instituto Del Radio, Azienda Ospedaliera Spedali Civili Di limitation encountered in planning these complex radiation treatments Brescia/ITALY, 9Dept Of Radiation Oncology, University Of Heidelberg/ that often require many optimization rounds and long optimization times. GERMANY, 10Dept Of Radiation Oncology, Zurich University Hospital, Therefore, the prescription dose goal of 5040cGy in 28 fractions is only Zurich/SWITZERLAND, 11Radiation Unit, Davidoff Center, Tel Aviv/ISRAEL achieved in approximately 33% of patients. Here we sought to develop a model that would predict the maximum safe radiation dose prescription for Background: Hemithoracic radiotherapy plays an important role in each patient. reducing locoregional relapse in malignant pleural mesothelioma patients following extrapleural pneumonectomy (EPP). Reports of patients treated Methods: We reviewed the radiation treatment plans of 56 MPM patients from 2003-2007 using intensity-modulated radiotherapy (IMRT) in with two intact lungs who were treated with definitive or adjuvant IMRT North America and Europe found significant toxicities from pneumonitis, after P/D. All patients were treated with coplanar 6MV beams using 6-9 including radiation-related fatalities that raised concerns for the safety of beam angles approximately equispaced over approximately 220 degrees IMRT. Following publication of more conservative constraints reports of to encompass the ipsilateral lung. The median dose was 4680cGy in 26 decreased toxicity have been published. However, since each institutional fractions (range 3960cGy – 5040cGy). Treatment plans were optimized to series presented results with small numbers, we sought to create an keep the mean lung dose (MLD) <21 Gy. Treatment planning parameters international mesothelioma interest group international database of IMRT toxicity for mesothelioma. including bilateral lung volumes, planning target volume (PTV), ipsilateral normal lung volume, mean total and individual lung doses, and prescription Methods: Institutions with experience in treating mesothelioma patients dose were recorded. The correlation between contralateral/ipsilateral lung with IMRT were surveyed to provide toxicity data on patients with volume ratio (CIVR), PTV/total lung volume ratio (PLVR) and prescription malignant pleural mesothelioma who received high-dose hemithoracic dose were analyzed. radiotherapy between January 2009 and December 2011. The incidence of CTCAE version 4 grade 3, 4 and 5 radiation pneumonitis was recorded Results: The median ipsilateral, contralateral and total mean lung doses together with demographic data, radiation lung dosimetry, timing of were 3941cGy, 534cGy and 2014cGy.The mean ipsilateral, contralateral chemotherapy and significant comorbidities. and total lung volumes were 1208cc, 1677cc and 2911cc, respectively. The median CIVR was 1.33. The only parameter in this study that correlated Results: Data was collected from 14 institutions in the USA, Canada, with achieving a higher prescription dose was CIVR (p= 0.005). France, Italy, Germany, Switzerland, Israel and Australia. All have used intensity-modulated radiotherapy to doses of 45-54 Gy in postpneumonectomy cases. In addition, data was collected on hemithoracic IMRT to patients with both lungs intact, after pleurectomy/decortication or non-surgical cases. 200 cases were reviewed with 9 cases of grade 3 or greater radiation pneumonitis. Detailed analysis of dose volume histogram data and clinical variables will be reviewed at the meeting. In addition, a comparison of tomotherapy and conventional IMRT delivered by linear accelerators will be presented.

Conclusion: With ongoing technological improvements in radiotherapy planning and equipment and tighter dose constraints, the incidence of serious radiation pneumonitis following high-dose hemithoracic IMRT has fallen in all institutions for patients treated after 2008. Further studies of selected non-EPP patients whose mesotheliomas are localized within one hemithorax should be undertaken.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 128 Conclusion: A higher ratio of contralateral/ipsilateral lung volume is an Conclusion: In the era of technological advances, HT offers a quite well important treatment planning parameter associated with achieving higher tolerated technique in treating MPM after EPP. The results are promising radiation prescription dose. Further investigation is needed to determine if but survival rates remain low for MPM patients completing multimodality other factors are significantly associated with a higher prescription dose. treatment.

Disclosure: No significant relationships. Disclosure: No significant relationships.

POSTER SESSION 3 September 13, 2012 11:30-12:30 POSTER SESSION 3 September 13, 2012 11:30-12:30

P3.29: HELICAL TOMOTHERAPY IN THE ADJUVANT TREATMENT P3.30: COMPARISON OF TWO DIFFERENT METHODS TO DELIVER OF RESECTED MALIGNANT PLEURAL MESOTHELIOMA: CLINICAL PLEURAL INTENSITY MODULATED RADIATION THERAPY IN OUTCOME AND TOXICITY. MALIGNANT PLEURAL MESOTHELIOMA

Joelle Helou1, Karen C. Colmou2, Alma Sylvestre1, Loic Campion2, Sophia Vishruta Dumane1, Andreas Rimner2, Ellen Yorke3, Abraham J. Zefkili3, Malika Amessis3, Pierre Bonnette4, Christian Perigaud5, Marc A. Wu2, Kenneth E. Rosenzweig4 Mahe2, Philippe Giraud1, Jacques Raphael6 1Department Of Radiation Oncology, Mount Sinai School Of Medicine, / poster sessions | september 13 1Radiation Oncology, Hopital Europeen Georges Pompidou, Paris/ NY/UNITED STATES OF AMERICA, 2Department Of Radiation Oncology, FRANCE, 2Radiation Oncology, Centre René Gauducheau, Nantes/ Memorial Sloan-Kettering Cancer Center, New York/UNITED STATES OF FRANCE, 3Radiation Oncology, Curie Institute/FRANCE, 4Chirurgie AMERICA, 3Department Of Medical Physics, Memorial Sloan-Kettering Thoracique Et Cardio-Vasculaire, Hopital Foch/FRANCE, 5Thoracic Surgery, Cancer Center, New York/NY/UNITED STATES OF AMERICA, 4Department Department Of Thoracic Surgery, University Hospital, Nantes, France, Of Radiation Oncology, Mount Sinai School Of Medicine, New York/NY/ Nantes/FRANCE, 6Thoracic Group, Inserm U981, Institut Gustave Roussy, UNITED STATES OF AMERICA Villejuif/FRANCE Background: The treatment of malignant pleural mesothelioma after Background: Malignant Pleural Mesothelioma (MPM) is an aggressive pleurectomy/decortication with radiation therapy remains a challenge tumour with poor prognosis. The incidence of local and distant recurrences due to the risk of pulmonary and cardiac toxicity. Recently, our group and after surgery remains high. Although a multimodality approach seems others have investigated the use of pleural intensity modulated radiation promising in reducing the incidence of local relapse, it is still limited by the therapy (IMRT) to treat patients with two intact lungs safely. IMRT can be high rate of toxicity. Our study aims to evaluate the outcome of patients delivered either by static field or via arc therapy. This study compares these with MPM treated by adjuvant Helical Tomotherapy (HT). two techniques to evaluate their potential radiation dosimetric advantage.

Methods: Between June 2007 and May 2011, 29 MPM patients received Methods: We compared plans for 4 left-sided and 3 right-sided cases. adjuvant radiotherapy by HT. The median age was 63 years (34-72) and the Plans used clinically approved planning target volumes (PTVs) and sex ratio (M/F) was 2.2. Twenty four patients had an epithelioid type MPM. critical organ contours. Static IMRT plans employed 7-8 6 MV photon Stage 2 and 3 diseases were observed in 12 and 17 patients respectively. beam directions over a 215º range centered on the ipsilateral lung. Arc Extra pleural pneumonectomy was performed in 25 patients. Thirteen therapy plans used 4 full arcs with avoidance sectors to better spare the patients received Platinum agent/antifolate regimen in the neoadjuvant contralateral structures. Prescription dose per fraction was 180 cGy. The setting. The median dose in the cavity of pneumonectomy was 50 Gy at prescription dose ranged from 4500 cGy to 5040 cGy so as to keep the 2 Gy / fraction. Event free survival (EFS) and overall survival (OS) were normal tissue complication probability (NTCP) of the combined lungs to ≤ calculated, adverse events were assessed by the RTOG toxicity grading 25%, our typical threshold. Planning objectives were: Lyman model NTCP scale. for both lungs < 25%; contralateral lung, mean dose < 8 Gy; heart, V30 Gy < 50%, mean < 30 Gy; each kidney, V18 Gy < 33%; liver _ not _ GTV, international mesothelioma interest group Results: Median follow up was 2.3 years. The most encountered acute side mean < 30 Gy, V30 Gy < 50%; stomach _ not _ PTV, mean < 30 Gy; effect was pulmonary grade 1-2 toxicity in 19 patients and grade 1-2 upper cord maximum < 45 Gy; bowel maximum < 55 Gy, D05 < 45 Gy; PTV D95 gastro-intestinal tract toxicity and/or dysphagia in 15 patients. Grade 3 ≥ 94%, V95 ≥ 94%, D05 ≤ 115%. Dose calculation was done with the AAA pneumonitis and dysphagia were observed in 1 and 2 patients respectively. algorithm. Contralteral lung V5 Gy, ipsilateral lung V30 Gy, V40 Gy and 2 cases of Grade 5 pneumonitis were suspected before 6 months following mean dose were also noted. RT. Late Grade 3-4 side effects were found in 4 patients. 19 patients had locoregional or distant relapse, 2 patients died of Grade 5 adverse events Results: Arc therapy lowered the average total lung NTCP from 16.9% to and 3 patients died from other causes. The median Event Free Survival 13% (p=0.03). The heart V30 decreased from 33.5% to 27.7% (p=0.016) (EFS) and Overall Survival (OS) were 1 and 1.6 y respectively. Fifty percent with arc therapy and the mean stomach dose decreased from 17 Gy to 14.7 of patients were event-free at 1 year and 22 % at 2 years. The overall Gy (p=0.03). The number of beams used and monitor units (a measure of survival at 1 and 2 years were 65% and 36% respectively. linear accelerator output) also decreased significantly with the use of arc therapy.

Conclusion: In this comparison of IMRT delivery either via static beams or arc therapy, we found a significant advantage in the use arc therapy to lower dose to the heart and lungs. Additionally, the use of arc therapy decreases the number of treatment beams and monitor units which decreases treatment time and increases patient comfort and compliance.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 129 Poster Session 4 Diagnostic and Predictive Biomarkers September 13, 2012 16:00-17:00

POSTER SESSION 4 September 13, 2012 16:00-17:00 POSTER SESSION 4 September 13, 2012 16:00-17:00

P4.01: CD26 OVEREXPRESSION IS ASSOCIATED WITH P4.02: HIGH DEFINITION CIRCULATING TUMOR CELLS (HD-CTC) PROLONGED SURVIVAL AND ENHANCED CHEMOSENSITIVITY IN IN MESOTHELIOMA. MALIGNANT PLEURAL MESOTHELIOMA 1 2 3 4 Lyudmila Bazhenova , Madelyn Luttgen , Dena Marrinucci , M Baughn , poster sessions | september 13 Keisuke Aoe1, Vishwa J. Amatya2, Nobukazu Fujimoto3, Kei Ohnuma4, Saskia Boisot4, Jorge Nieva5, Kelly Bethel4, Peter Kuhn2 Osamu Hosono5, Akio Hiraki6, Masanori Fujii7, Taketo Yamada8, Nam 1Medical Oncology, Ucsd Moores Cancer Center, La Jolla/UNITED STATES H. Dang9, Yukio Takeshima2, Kouki Inai2, Takumi Kishimoto10, Chikao OF AMERICA, 2The Scripps Research Institute, La Jolla/UNITED STATES OF Morimoto4 AMERICA, 3Epic Sciences, La Jolla/UNITED STATES OF AMERICA, 4Scrips 1Medical Oncology, Yamaguchi-Ube Medical Center, Ube, Yamaguchi/ Clinic, La Jolla/UNITED STATES OF AMERICA, 5Billings Clinic, Billings/ JAPAN, 2Pathology, Institute Of Biomedical & Health Sciences Hiroshima UNITED STATES OF AMERICA University, Hiroshima/JAPAN, 3Respiratory Medicine, Okayama Rosai Hospital/JAPAN, 4Rheumatology And Allergy, Institute Of Medical Science, Background: Circulating tumor cells (CTCs) are emerging as a valuable University Of Tokyo/JAPAN, 5Clinical Immunology, Institute Of Medical tool for monitoring cancer patient prognosis and response to treatment in Science, University Of Tokyo/JAPAN, 6Mizushima Daiichi Hospital/ some solid tumors. Commercially available CTC assay identifies CTCs via JAPAN, 7Respiratory Medicine, Japanese Red Cross Kobe Hospital/ EPCAM dependent immunomagnetic-enrichment, which may not detect JAPAN, 8Pathology, Keio Univarsity/JAPAN,9Hematology/Oncology, certain subsets of cells. We report a cohort of mesothelioma patients University Of Florida/UNITED STATES OF AMERICA, 10Respiratory Medicine, in whom CTCs were measured using a cytometric, enrichment free Okayama Rosai Hospital, Okayama/JAPAN immunofluorescent protocol and correlated with outcomes.

Background: Malignant pleural mesothelioma (MPM) is an aggressive Methods: Blood samples for CTC analysis were collected from 9 and therapy-resistant neoplasm arising from the pleural mesothelial mesothelioma patients at baseline, 3 weeks, 3 months, 6 months, 9 cells. There are no established indicators to predict responsiveness to months, and 1 year. HD-CTCs were identified via immunofluorescence and chemotherapeutic treatment for MPM. cytometric, enrichment free analysis. We also reviewed primary tumor specimens and correlated cytomorphologic appearance of the HD-CTCs to Methods: Our present study involving 79 MPM patients demonstrated that their corresponding primary tumor. 73.4% of MPM expressed CD26 on cell membrane. Results: A total of 9 consecutive chemotherapy naïve mesothelioma Results: The majority of epithelioid and biphasic type of MPM expressed patients were included in the analysis. All had non-sarcomatoid histology. CD26 on the membrane, whereas sarcomatoid type demonstrated a lack Median follow up was 18.8 months (range 4.6 to 24.4). Median survival is of CD26 surface expression. Although sarcomatoid type was associated 19.6 months (range 4.6 to 24.3). At some point during the disease course international mesothelioma interest group with poor prognosis (p<0.0001), no significant relationship between CD26 CTCs were identified in 6 of 8 (67%) patients (range 0-566 CTC/ml, mean expression and survival was observed (p=0.1384). On the other hand, 17 CTC/ml). (see table) At the time of the diagnosis, CTCs were present in 5 the response rate to chemotherapy was marginally associated with CD26 out of 9 patients but only 1 of those patients had distant metastases. Three expression (P=0.053), with higher level of CD26 expression more likely to of 9 patients were alive at the time of data analysis. Using CTC/ml cut off be linked to better response to chemotherapy. Moreover, CD26 expression of 2 CTCs/ml, patients can be divided into ‘remain favorable’ (baseline was a significant factor associated with improved survival in chemotherapy draw <2, last draw <2), ‘convert to favorable’ (baseline draw >2, last draw patients (MST, 18.6 vs 10.7 months, P=0.0083). <2), remain unfavorable (baseline draw >2, last draw >2), and convert to Furthermore, CD26 expression was significantly associated with better unfavorable (baseline draw <2, last draw >2). Median survival is 604 days prognosis in patients with non-pemetrexed (PEM) regimens (MST, 14.2 for ‘remain favorable’/’convert to favorable’ group and 475 days for the vs 7.4 months, P=0.0042), while there was no significant association ‘remain unfavorable’/’convert to unfavorable’ group. Presence of CTCs at between CD26 expression and survival time for patients with PEM regimens the time of diagnosis was not predictive of survival. (P=0.1225). Our in vitro and microarray studies showed that mesothelioma cells expressing high CD26 display high proliferative activity, and CD26 expression is closely linked to cell proliferation, cell cycle regulation, apoptosis, and chemotherapy resistance.

Conclusion: Our results strongly suggest that CD26 is a clinically significant biomarker for predicting response to chemotherapy for MPM.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 130 received CT in 1st line and 34.7% in 2nd line. We found a significant CTC counts association of calretinin expression with epithelial histology (p= 0.030) and PS 1 (p= 0.05) and no association with clinical stage (p= 0.23). WT1 expression was associated with epithelial histology (p= 0.010), but no Patient Stage # Range Median Mean % Survival association with PS and clinical stage (p<0.05) was detected. After a number draws (CTC/ (CTC/ (CTC/ draws (mos) median follow up of 9.2 months the median survival (OS) was 15.2 months mL) mL) mL) positive for the entire cohort. We found a significant increase in OS in patients with epithelial subtype (23.4 vs 5.0 months in epithelial vs no-epithelial, 1 T4N2M1 7 0-0.9 0 0.12 14.2 20.4 p<0.001), PS1 (14.7 m vs 2.2 months in PS 1 vs PS 2, p=0.036), NLR ≤5 2 T4N0M0 7 0 -7.9 0 1.12 14.2 23.1 (26.5 vs 13.4 months, p=0.025) and patients who received 2nd line CT (8.9 and 26.4 months for p second line, p=0.05). In the IHC markers analysis 3 T3N0M0 5 0-0 0 0 0 24.7 we found a significant increase in OS for p with WT1 positive expression 4 T2N1M1 2 2.6-108 55.5 55.5 100 4.6 (16.4 m vs 2.3 m, p= 0.013), but not differences for calretinin expression (16.6 m vs 5.0 months, p=0.37). In the multivariate analysis epithelial 5 T2N2M0 7 0-16.5 1.3 5.08 86% 18.8 histology and WT1remained as significant prognostic factors for survival 6 T3N0M0 5 0-31.8 1 7.08 60% 11.5 (HR 22.8; 95%CI,2.7-190, p=0.004 and HR 9.5; 95%CI 1.7-52.3, p=0.010 respectively).

7 T2N2M0 6 0-566.4 0.5 95.3 50% Alive poster sessions | september 13 (21.8) Conclusion: In our series of 52 MPM patients, epithelial histology, PS, NLR, 8 T2N0M0 3 0-2.1 1 1.03 66% Alive second line CT and WT1 expression are significant prognostic factors for (17.1) survival. The role of WT1 assessment is worth of prospective validation in 9 T2N0M0 1 0 0 0 0 Alive future studies on MPM. (8.6) Disclosure: No significant relationships.

Conclusion: This small case series shows that CTCs can be effectively enumerated in mesothelioma patients. CTC presence in mesothelioma patients does not signify presence of metastatic disease or predict POSTER SESSION 4 September 13, 2012 16:00-17:00 development of metastases. There appears to be a separation in outcomes based on CTC kinetics. However, due to the small sample size further P4.04: SERUM CA-125 IS ASSOCIATED WITH WORSE SURVIVAL studies are warranted to further clarify this association. IN MALIGNANT PLEURAL MESOTHELIOMA: RETROSPECTIVE ANALYSIS IN A SINGLE CENTER SERIES. Disclosure: Epic Sciences Susana Cedres1, Alex Martinez-Marti2, Pablo Martinez2, Davis Torrejon1, Maria Angeles Montero2, Enriqueta Felip2 1Medical Oncology, Vall D´hebron University Hospital, Barcelona/ POSTER SESSION 4 September 13, 2012 16:00-17:00 SPAIN, 2Medical Oncology Department, Vall D´hebron University Hospital/ SPAIN P4.03: EXPRESSION OF WILMS´TUMOUR GENE (WT1) IS ASSOCIATED WITH SURVIVAL IN MALIGNANT PLEURAL Background: Malignant pleural mesothelioma (MPM) is an aggressive MESOTHELIOMA: RETROSPECTIVE ANALYSIS IN A SINGLE and difficult to treat tumor. Serum tumor markers have been used in CENTER SERIES mesothelioma as diagnostic tools complementary to pathological findings. However, the prognostic value of serum tumor markers in MPM has not Susana Cedres1, Pablo Martinez1, Alex Martinez-Marti1, Maria Angeles been evaluated. We investigated the overall survival (OS) according to international mesothelioma interest group Montero2, Davis Torrejon1, Enriqueta Felip1 serum tumor markers in a series of patients diagnosed of MPM in our 1Medical Oncology, Vall D´hebron University Hospital, Barcelona/ institution SPAIN, 2Pathology, Vall D´hebron University Hospital, Barcelona/SPAIN Methods: Fifty two patients diagnosed of MPM in Vall d´Hebron Background: Calretinin and Wilms´tumour gene (WT1) are mesothelial University Hospital between November 2002 and September 2011 were markers routinely used to confirm the diagnosis of malignant retrospectively reviewed. Baseline characteristics analyzed were age, pleural mesothelioma (MPM). Recently, calretinin score assessed by performance status (PS), neutrophil to lymphocyte ratio (NLR), clinical immunohistochemistry (IHC) was implicated with poor prognosis in MPM. stage, histology, and chemotherapy treatment (CT). Serum tumor markers We investigated the prognostic value of calretinin and WT1 expression analyzed were CEA, CYFRA 21-1, CA-125, enolase and Ca15.3. All tumor in predicting survival in a series of patients (p) diagnosed of MPM in our markers were performed during the study of suspected malignancy. institution Survival data were calculated by the Kaplan-Meier method and Cox regression was used to evaluate the prognostic value of the variables. Methods: Fifty two patients diagnosed of MPM in Vall d´Hebron University Hospital between November 2002 and September 2011 were Results: Patient’s characteristics: median age 68 years (31-88 years), retrospectively reviewed. Potential prognostic factors analyzed were age, males 75.5%, PS 1: 67.3%, asbestos exposure 53.1%, clinical stage III: performance status (PS), neutrophil to lymphocyte ratio (NLR), clinical 55.1%, epithelial subtype 71.4% and NLR>5 in 44.9% of all patients. CEA stage, histology, calretinin and WT1 expression, and chemotherapy was available in 35 p; CYFRA21-1 in 24 p; CA-125 in 31p; enolase in 21p; treatment (CT). Calretinin and WT1 were stained for IHC analysis in and CA15.3 in 16 p. We did not found association for any tumor marker with formalin-fixed, paraffin-embedded sections and were considered positive histology, stage, gender, smoking history, pleural effusion and asbestos cases with >1% of tumor cells stained. Survival data were calculated by exposure. We found frequent elevation above the upper limit of the the Kaplan-Meier method and Cox regression was used to evaluate the serum tumor markers at diagnosis in MPM. CEA was elevated in 5.8% p; prognostic value of the variables. CYFRA2-1- in 62.5% p; CEA-125 in 51.6%p; enolase in 19% p and Ca 15.3 in 62.5% of patients. After a median follow up of 9.2 months the median Results: Patient’s characteristics: median age 68 years (31-88 years), survival was 15.2 months for the entire cohort. We found a significant males 75.5%, PS 1: 67.3%, asbestos exposure 53.1%, clinical stage III: increase in OS in patients with epithelial subtype (23.4 vs 5.0 months in 55.1%, epithelial subtype 71.4% and NLR>5 in 44.9% of all patients. no-epithelial, p<0.001), PS1 (14.7 vs 2.2 months in PS 2, p=0.036), NLR Calretinin IHC expression was available in 47 p and was positive in 41 p ≤5 (26.5 vs 13.4 months, p=0.025) and patients who received 2nd line CT (83.7%). WT-1 IHC expression was available in 32 p and was positive in 25 (8.9 and 26.4 months for p second line, p=0.05). In the analysis of tumor p (78.1%). All patients were considered initially unresectable and 71.4% markers we found that CA-125 was the only tumor marker significantly

iMig2012.org • Abstract Book 131 associated with survival. Patients with CA-125 elevated had a median POSTER SESSION 4 September 13, 2012 16:00-17:00 survival of 2.1months vs 18.9 months for patients CA-125 in normal value (p=0.04). No differences in survival were detected with the CEA, P4.06: CCL2, GALECTIN-3 AND SMRP COMBINATION IMPROVES CYFRA21-1, enolase and Ca15.3 (p>0.05). In the multivariate analysis only THE DIAGNOSIS OF MESOTHELIOMA IN PLEURAL EFFUSIONS epithelial histology remained as significant prognostic factors for survival (HR 7.05; 95%CI,1.93-25.73, p=0.003). Marc Gregoire1, Christophe Blanquart1, Fabien Guegnon1, Jean-michel Nguyen2, Laurent Cellerin3, Christine Sagan4, Christian Perigaud3, Conclusion: In our series of 52 MPM patients, epithelial histology, PS, NLR Arnaud Scherpereel5 and second line CT are significant prognostic factor for survival. We found 1Inserm-892/Cnrs-6299, Institute Of Therapeutic Research, Nantes/ that basal level of CA-125 is a significant prognostic factor for survival in FRANCE, 2Seb-Pimesp, Nantes Hospital, Nantes/FRANCE, 3Service mesothelioma. Oncologie Médicale Thoracique Et Digestive, Hospital Laennec, Nantes/ FRANCE, 4Service D’Anatomie Pathologique, Nantes Hospital, Nantes/ Disclosure: No significant relationships. FRANCE, 5Service De Pneumologie Et Oncologie Thoracique, Lille Hospital, Lille/FRANCE

Background: Malignant Pleural Mesothelioma (MPM) is a highly aggressive POSTER SESSION 4 September 13, 2012 16:00-17:00 tumor. Diagnosis, which is sometimes difficult, is usually established at

an advance stage of the disease when therapies are of limited efficiency. poster sessions | september 13 P4.05: PLEURAL HYALURONIC ACID IS A USEFUL BIOMARKER FOR Thus, one main challenge for this disease is the development of new, MALIGNANT MESOTHELIOMA early and highly-reliable diagnostic markers. The aim of this work was to compare the diagnostic value of three new soluble markers (the chemokine Jenette Creaney1, Amanda Segal2, Ian Dick1, Arthur W. Musk3, Bruce CCL2, galectin-3 (LGALS3) and the secretory leucocyte peptidase inhibitor W.S. Robinson4 SLPI) with soluble mesothelin-related peptides (SMRP) and to evaluate the 1School Of Medicine And Pharmacology, University Of Western Australia, diagnostic performance of markers combinations. Nedlands/AUSTRALIA, 2Pathwest, Nedlands/AUSTRALIA, 3Respiratory Medicine, Sir Charles Gairdner Hospital, Nedlands/WA/AUSTRALIA, 4School Methods: 101 pleural fluids from patients with suspicion of MPM were Of Medicine And Pharmacology, University Of Western Australia, Perth/ collected between 1998 and 2010. The levels of the different biomarkers WA/AUSTRALIA were measured using commercial Enzyme-linked immunosorbent assays.

Background: The use of biomarkers in pleural effusions has not been Results: We found that SMRP concentration was significantly higher in clinically accepted. Whilst good discrimination between malignant and pleural effusions from patients with MPM (n=61) than in subjects with benign effusions has been demonstrated using various biomarkers, adenocarcinoma (ADCA, n=25) or with benign pleural effusions (BPE, their ability to assign phenotypes to particular malignancies has been n=15) [median (interquartile range)=33.8 nM (13.85–84.15) versus 3.4 limited. Hyaluronic acid (HA) is a glycoaminoglycan previously shown to nM (1.2-7.7) and 2.5 nM (0-7.1), respectively, P< 0.0001]. Likewise, SLPI be elevated in malignant mesothelioma (MM) effusions and may be useful concentration was higher in MPM than in ADCA and BPE [228.2 ng/ for discriminating between MM and other pleural metastatic cancers. In mL (113.6-409.8) versus 101.5 ng/mL (72.9-154.5) (P< 0.001) and 91.1 this study we compared HA and mesothelin in serum and pleural effusions ng/ml (68-158.1) (P< 0.05). ROC curves analysis revealed that SMRP to investigated the ability to distinguish MM from lung adenocarinoma (AUC=0.9059), CCL2 (AUC=0.7912), Galectin-3 (AUC=0.7584) and SLPI metastatic to the pleura (AUC=0.7219) were potential interesting biomarkers to differentiate MPM patients from patients with BPE or ADCA. Interestingly, we showed Methods: Serum and pleural effusate was collected from 92 MM patients, that combination of SMRP, CCL2 and Galectin-3 greatly improved MPM 24 lung cancer (LC) patients and 43 patients with benign pleural effusions. diagnosis (AUC=0.9680). HA was measured using the HA Binding assay (Corgenix Inc). Mesothelin was measured using the MESOMARK TM assay (Fujirebio). A pleural effusate Conclusion: The measurement of SLPI, CCL2 and Galectin-3 levels in international mesothelioma interest group was defined as positive when HA concentrations were above the mean of pleural fluids allowed the diagnosis of MPM with good sensitivity and the benign control group by two standard deviations, i.e. above 50 mg/ml. specificity. However, SMRP remained the best single soluble marker The previously defined threshold for mesothelin in effusions of 20nM was available. The combination of SMRP, CCL2 and Galectin-3 evaluation in chosen to dichotomise samples. pleural fluid appears as a promising panel of biomarkers for a reliable diagnosis of MPM. Results: There was no significant difference in the serum HA level between the groups. HA concentrations were approximately 1000 fold Disclosure: No significant relationships. higher in the effusion than serum of MM patients. In pleural effusions, HA concentrations were significantly higher in MM compared to lung cancer (p<0.001) and effusions of benign aetiology (p<0.001). With less than 5% of lung cancer and benign effusion cases being HA positive compared POSTER SESSION 4 September 13, 2012 16:00-17:00 to approximately 60% of MM cases. There was a significant correlation between pleural effusion mesothelin and HA concentrations. However, P4.07: A RETROSPECTIVE MULTICENTER EVALUATION OF there was no significant difference between the diagnostic accuracy of PRETREATMENT SERUM C-REACTIVE PROTEIN LEVELS AS the two biomarkers when the area under the receiver operator curves was PROGNOSTIC AND PREDICTIVE MARKER IN PATIENTS WITH compared. MALIGNANT PLEURAL MESOTHELIOMA

Conclusion: These preliminary data demonstrate biomarkers have the Mir A. Hoda1, Bahil Ghanim1, Maximilian P. Winter1, Thomas Klikovits1, potential to not only discriminate between malignant and benign effusions Balazs Hegedus1, Balazs Dome1, Madleine Arns2, Peter Schenk2, but also indicate malignant phenotype. Wolfgang Pohl3, Christoph Zielinski4, Martin Filipits5, Walter Klepetko1, Walter Berger5 Disclosure: No significant relationships. 1Division Of Thoracic Surgery, Medical University Of Vienna, Vienna/AUSTRIA, 2Pneumology, Lkh Grimmenstein-Hochegg/ AUSTRIA, 3Pneumology, Kh Hietzing/AUSTRIA, 4Division Of Clinical Oncology, Medical University Of Vienna/AUSTRIA, 5Institute Of Cancer Research, Medical University Of Vienna/AUSTRIA

Background: Malignant Pleural Mesothelioma (MPM) is an aggressive disease with poor prognosis. Therapeutic decision is challenging and

iMig2012.org • Abstract Book 132 biomarkers for better treatment stratification are needed. The main aim Disclosure: No significant relationships. of this study was to evaluate the prognostic and predictive relevance of pretreatment serum C-reactive protein (CRP) in malignant pleural mesothelioma (MPM) patients. POSTER SESSION 4 September 13, 2012 16:00-17:00 Methods: Clinical data were retrospectively collected from 115 patients with histologically proven MPM. Patients with evidence for infectious P4.09: PDGF-D/PDGF-ββ RECEPTOR-REGULATED CHEMOTAXIS disease were excluded. The association between CRP levels and survival OF MALIGNANT MESOTHELIOMA CELLS was analyzed using Kaplan-Meier method and Cox models adjusted for clinico-pathological factors. Asuka Okada1, Takahiro Yaguchi2, Hitomi Kamiya1, Miki Honda1, Eriko Masachika1, Hisaya Okuwa1, Taiichiro Otsuki1, Koji Mikami1, Results: Median pretreatment CRP of all patients was 1.19 mg/dl (range: Yoshitaka Nogi1, Risa Maeda1, Takayuki Terada1, Noriko Hirayama1, 0.00-22.62 mg/dl). Patients with elevated CRP (≥1mg/dl; n=62, 53.9%) Shusai Yamada1, Kunihiro Tamura1, Chiharu Tabata1, Kazuya Fukuoka1, had a significantly shorter overall survival (OS) compared to those with Tomoyuki Nishizaki2, Takashi Nakano1 normal CRP (HR 2.81, 95% CI 1.82-4.33; p<0.001). In multivariate analyses, 1Division Of Respiratory Medicine, Department Of Internal Medicine, elevated CRP was confirmed as independent prognosticator in MPM (HR Hyogo College Of Medicine, Nishinomiya/JAPAN, 2Division Of Bioformation, 2.07, 95% CI 1.23-3.46; p=0.01). A significant interaction between CRP Department Of Physiology, Hyogo College Of Medicine, Nishinomiya/JAPAN and treatment modality (p<0.001) was observed. Among patients with poster sessions | september 13 normal CRP, radical tumor resection within multimodality protocols (MMP) Background: Our earlier study suggested that platelet-derived growth was associated with prolonged OS when compared to protocols without factor (PDGF)-ββ receptor regulates chemotaxis of human malignant surgery (HR 7.26, 95% CI 3.40-15.49; p<0.001). Among patients with mesothelioma cells such as MSTO-211H, NCIH-2052, NCIH-2452, and elevated CRP, no survival benefit was achieved by radical surgery within NCIH-28 cells, but not non-malignant Met5A cells. The present study MMP (HR 0.911, 95% CI 0.53-1.58; p=0.74). was designed to gain further insight into the PDGF-ββ receptor signals underlying the chemotaxis. Conclusion: Our results suggest that multimodality regimens including radical resection increase survival selectively in MPM patients with normal Methods: PDGF-D secreted from cells, activation of Akt and ERK, and cell pretreatment serum CRP levels. migration were monitored for cells with and without knocking-down PDGF- ββ receptor. Disclosure: No significant relationships. Results: FBS significantly stimulated PDGF-D secretion from malignant mesothelioma cells, but not Met5A cells. PDGF-D activated Akt and ERK in both the non-malignant and malignant cells. PDGF-D significantly POSTER SESSION 4 September 13, 2012 16:00-17:00 facilitated migration of malignant mesothelioma cells, but not Met5A cells, with the extent varying among the cell types. The facilitatory action of P4.08: PERIPHERAL T CELL ACTIVITY PREDICTS SURVIVAL IN PDGF-D was clearly prevented by knocking-down PDGF-ββ receptor or PATIENTS WITH MALIGNANT MESOTHELIOMA AND NON-SMALL inhibitors of PI3 kinase, PDK1, Akt, Rac1, ROCK, and MEK. CELL LUNG CANCER Conclusion: The results of the present study indicate that PDGF-D Melanie J. Mccoy, Anna K. Nowak, Robbert G. Van Der Most, Alison M. promotes malignant mesothelioma cell chemotaxis through PDGF-ββ Mcdonnell, Ian M. Dick, Richard Lake receptor signaling pathways along a PI3 kinase/PDK1/Akt/Rac1/ROCK axis School Of Medicine And Pharmacology, National Centre For Asbestos and relevant to ERK activation. Related Diseases, University Of Western Australia, Perth/WA/AUSTRALIA Disclosure: No significant relationships. Background: The importance of generating an anti-tumor immune international mesothelioma interest group response is manifest in the way that the balance of intratumoral T cell subsets reflects clinical outcome. Tumor-infiltration by CD8+ T cells and regulatory T cells (Treg) is associated with improved and reduced POSTER SESSION 4 September 13, 2012 16:00-17:00 survival respectively in many cancer types. However, little is known of the prognostic value of immunological parameters measured in peripheral P4.10: PLEURAL FLUID CELL-FREE DNA ABSOLUTE TELOMERE blood. This study aimed to characterize the relationship between the LENGTH IN MALIGNANT AND BENIGN EFFUSIONS proportion/phenotype of peripheral blood T cell subsets and survival following palliative chemotherapy in patients with malignant mesothelioma Krishna B. Sriram1, Vandana Relan1, Belinda Clarke2, Edwina Duhig2, (MM) or advanced non-small cell lung cancer (NSCLC). Morgan Windsor3, Kevin Matar3, Rishendran Naidoo3, Linda Passmore1, Elizabeth Mccaul1, Deborah Courtney1, Ian A. Yang1, Rayleen V. Methods: Blood samples were collected from 43 patients with MM Bowman1, Kwun M. Fong1 or advanced NSCLC prior to commencement of platinum-doublet 1Department Of Thoracic Medicine, The Prince Charles Hospital, Brisbane/ chemotherapy. The proportions of activated, proliferating and tumor- QLD/AUSTRALIA, 2Department Of Anatomical Pathology, The Prince Charles associated antigen (TAA)-specific T cell subsets were assessed by multi- Hospital/AUSTRALIA, 3Department Of Thoracic Surgery, The Prince Charles parameter flow cytometry. Hospital/QLD/AUSTRALIA

Results: Patients had a higher proportion of peripheral Treg, proliferating Background: Alterations in telomere length (increased and shortened) in CD8+ T cells and activated effector CD8+ and CD4+ T cells compared to peripheral blood leucocytes and tumour tissue have been associated with healthy controls of a similar age. These differences did not correlate with malignancies, including malignancies which metastasise to the pleura, markers of systemic inflammation. TAA-specific CD8+ T cells were detected presenting as malignant pleural effusions. The aim of this study was to in 10% of patients. Higher proportions of Treg and proliferating CD8+ T quantitatively measure absolute telomere length in pleural fluid cell-free cells prior to chemotherapy were both associated with poor survival in DNA in subjects with malignant compared with benign pleural effusions. univariate analyses (hazard ratio [HR] = 3.81; 95% CI, 1.69 to 8.57; p = 0.001 and HR = 2.86; 95% CI, 1.26 to 6.50; p = 0.012 respectively). CD8+ T Methods: We studied pleural fluid samples from 96 consecutive subjects cell proliferation was independently predictive of reduced survival in (68 malignant and 28 benign effusions). Absolute telomere length multivariate analysis (HR = 3.14; 95% CI, 1.29 to 7.65; p = 0.012). was measured in pleural fluid cell-free DNA (cfDNA) using a modified quantitative PCR method. Median absolute telomere length was compared Conclusion: Peripheral T cell activity may be a useful prognostic marker in in the pleural fluid cfDNA between subjects with malignant and benign patients with thoracic malignancies planned for palliative chemotherapy. effusions.

iMig2012.org • Abstract Book 133 Results: The median age of the MPE and benign effusions subjects was 69 value below threshold. This algorithm remained consistent in a narrow years and 74 years respectively. Male subjects comprised 53% of the MPE range in 38 out of 42 individuals with at least 3 visits over a minimum 2 group compared to 75% in the benign effusion group. Sixty-three percent year period. of MPE subjects were current or former smokers compared to 75% of subjects with benign effusions. The most common aetiologies of MPE were Conclusion: An algorithm combining both biomarkers remained consistent lung cancer (n=27), mesothelioma (n=23) and breast cancer (n=7) while the over time in a clinical screening setting. Considering the rapid progression most common aetiologies for benign effusions were inflammatory pleuritis of mesothelioma, our early screening program using low dose computed (n=11), asbestos related effusions (n=6) and parapneumonic effusions tomography can be refined with the use of tumour biomarkers. (n=5). The median absolute telomere length measured in pleural fluid cfDNA was not significantly different in subjects with malignant pleural Disclosure: No significant relationships. effusions compared to subjects with benign effusions (9.7kb per diploid genome vs. 8.5kb per diploid genome, p=0.488). There were no significant differences in the median telomere lengths in subjects with malignant pleural mesothelioma (10.3kb vs. 8.5kb, p=0.385), lung cancer (8.5kb vs. 8.5kb, p=0.390) or other cancers (4.7kb vs. 8.5kb, p=0.384) compared to those with benign pleural effusions. When we categorized the pleural fluid cfDNA absolute telomere length into quartiles based on the telomere length distribution of the controls, with the first (shortest) quartile poster sessions | september 13 being used as the reference category, the age-adjusted OR for MPE was essentially equal in the 2nd (0.92, 95% CI 0.83-1.02; p=0.10), 3rd (1.01; 95% CI0.95 – 1.06, p=0.896) and 4th quartiles (1.01, 05% CI 0.95-1.07, p=0.76). In the study cohort, covariates were examined for relationships with absolute telomere length. There was no difference in pleural fluid cfDNA absolute telomere length for age <60 years compared to age > 60 years (8.0kb vs. 9.7kb, p=0.187), females compared to males (9.5kb vs. 9.9kb, p=0.548), cytology positive compared to cytology negative (8.99kb vs. 9.38kb, p=0.924). There was a trend for shorter telomeres in smokers compared with non-smokers, however the difference did not reach statistical significance (9.0kb vs. 12.3kb, p=0.076).

Conclusion: In this study we found that it was technically possible to measure absolute telomere length in pleural fluid cfDNA. However the pleural fluid cfDNA is most likely a mixture of various types of white blood cells, mesothelial and tumour cells. Further study is required to determine the absolute telomere length in the individual cells and this may be possible by initially identifying and sorting the individual cell groups using flow ctyometry.

Disclosure: No significant relationships.

POSTER SESSION 4 September 13, 2012 16:00-17:00 international mesothelioma interest group P4.11: SOLUBLE MESOTHELIN RELATED PROTEIN AND OSTEOPONTIN IN SCREENING FOR MALIGNANT PLEURAL MESOTHELIOMA

Katrina Rey-Mcintyre1, Masaki Anraku1, Licun Wu1, Tetsuzo Tagawa1, Zhihong Yun1, Brenda O’Sullivan2, Zhuo Chen2, Geoffrey Liu2, Demetris Patsios2, Lu Chen2, Wei Xu2, Ming Tsao2, Marc De Perrot3 1Latner Thoracic Surgery Research Laboratories, Toronto General Research Institute, Toronto/ON/CANADA, 2University Health Network, Toronto/ CANADA, 3Latner Thoracic Surgery Research Laboratories And Division Of Thoracic Surgery, Toronto/ON/CANADA

Background: We established a screening program for asbestos exposed individuals using low-dose computed tomography. Incorporating biomarkers could provide more effective screening. We determined the diagnostic performance of osteopontin combined with soluble mesothelin related protein using an algorithm and determined the variance of the algorithm values over several years.

Methods: Plasma was obtained from 67 patients with malignant pleural mesothelioma and 278 asbestos exposed controls from our screening program.

Results: Using an algorithm of both biomarkers, values were significantly higher in patients with malignant pleural mesothelioma compared to asbestos exposed controls in a test group and in a validation group. Using receiver operating characteristic curves, an area under curve of 0.88 was produced for the test group and 0.83 for the validation group. A total of 114 out of 118 individuals tested in the screening program had an algorithm

iMig2012.org • Abstract Book 134 Poster Session 4 Multimodality September 13, 2012 16:00-17:00

POSTER SESSION 4 September 13, 2012 16:00-17:00 POSTER SESSION 4 - September 13, 2012 16:00-17:00

P4.12: EXTRAPLEURAL PNEUMONECTOMY IN EPITELIOID P4.13: A CLINICAL STUDY OF 12 DIFFUSE MALIGNANT PLEURAL MALIGNANT PLEURAL MESOTHELIOMA: IS IT OBSELETE OR IS MESOTHELIOMA IN 70 YEARS OR OLDER THERE A NEED FOR A LARGER RANDOMIZED TRIAL?

Takuro Fushimi, Osamu Kawamata poster sessions | september 13 Onur Ermerak1, Adamu Issaka1, Barkin Eldem1, Hakan Ozalper1, Zeynep Surgery, Onomichi Municipal Hospital, Onomichi/JAPAN Bilgi1, Fulden Yumuk2, Hale Basak Ozkok3, Muzaffer Metintas4, Volkan Kara1, Hasan F. Batirel1 Background: Diffuse malignant pleural mesothelioma (MPM) is a highly 1Thoracic Surgery, Marmara University Hospital, Istanbul/TURKEY, 2Division aggressive tumor with very poor prognosis, but opitimal treatment has Of Medical Oncology, Department Of Internal Medicine, Marmara not been defined yet. Asbestos exposure is the main factor involved University Hospital, Istanbul/TURKEY, 3Department Of Radiation Oncology, in pathogenesis of MPM. Because of the long incubation time, it is Acibadem University, Istanbul/TURKEY, 4Department Of Pulmonary expected the number of elder patients will increase. We consider about Medicine, Osmangazi University Hospital, Istanbul/TURKEY chemotherapy for elderly MPM patients.

Background: The unfavorable survival figures in the only randomized Methods: Between 2000 and 2011, 24 MPM patients were definitive trial in malignant pleural mesothelioma (MPM) have sparked discussions diagnosed by thoracoscopic biopsy in our hospital, and we analyzed 12 of unnecessity for the application of extrapleural pneumonectomy (EPP). cases over 70 years when diagnosed. However, this conclusion could prevent a subgroup of patients who would benefit from EPP and multimodality treatment. We analyzed our data in Results: Of the 12 patients, 10 were men and 2 were women with the patients with epithelioid MPM. mean age 77.3 years old (7 were in 70’s, 5 were in 80’s). 8 were epithelial type, 2 were sarcomatoid type, and 2 were biphasic type in histology. 2 Methods: Patients who underwent EPP for the treatment of epitelioid were StageIb, 8 were StageII, 1 was StageIII, and 1 was StageIV according MPM in two hospitals between 2002-2011 were included in the study. to IMIG. 8 patients were treated with chemotherapy (6 epithelial, 2 EPP technique included en bloc resection of ipsilateral pleura, lung, biphasic). Of the 8, 2 were underwent extrapleural pnumonectomy (EPP). pericardium and diaphragm. Patients were referred for adjuvant 4 patients were treated symptomatic therapy. Of the 8 chemotherapy hemithoracic radiation and platinum based chemotherapy. All demographic patients, 3 were performed cisplatin(CDDP)+Pemtrexed, 2 were performed and patient data was recorded in a prospective database and statistical CDDP+Gemcitabine(GEM), 2 were performed GEM+Vinorelbine, and 1 was analysis was performed using Kaplan Meier survival curves and uni- and performed GEM alone. In chemotherapy group, 3 are alive (2 were received multivariate analysis for determination of prognostic factors. EPP, monitoring periods; 11.5, 22.3, 33.7 months) 5 were dead (OS; 18.1, 23.4, 28.3, 31.7, 34.6 months). In non-chemotherapy group, all were dead

Results: 40 patients underwent EPP during this period. 35 had epitelioid (OS; 3.8, 4.8, 6.6, 7.9 months). 3 patients those were performed GEM international mesothelioma interest group tumors (Average age 54 ± 8, 14 females). In-hospital mortality was 12,5% alone (over 6 months, include second line) shows long prognosis without (6% at University hospital). T stages (IMIG staging) were 1 (n=2), 2 (n=25), losing their Quality of Life (QOL). Their OS and monitoring time were 3 (n=7), 4 (n=1). 14 had extrapleural lymph node metastasis and 9 also had 23.4months (dead, GEM alone periods; 14 months), and 22.3, 33.7 months intraparenchymal lymph node metastasis. 25 patients (66%) completed (alive, GEM alone periods; 9.6, 15.6 months, sepalately). adjuvant chemoradiation. Overall median survival was 18,1 months (39% 2-year, 11% 5-year survival. Females had better overall median survival Conclusion: This analysis provides active treatments bring better prognosis when compared with males (25,7 vs 15,5 months respectively) although even for the elderly MPM patients. It was thought that GEM alone it did not reach significance (p=0,1). Females who completed adjuvant chemotherapy may keep their better QOL. chemoradiation (n=12) had a median survival of 31,8 months (22% 5-year survival). Median survival was not different in patients who had Disclosure: No significant relationships. extrapleural lymph node metastasis and who did not (16,9 vs 20,1 months, p=0,22).

Conclusion: There is a subgroup of patients who benefit from EPP and POSTER SESSION 4 September 13, 2012 16:00-17:00 adjuvant chemoradiation with a 5-year survival rate over 20%. EPP allows us to accurately stage patients and provide data that may be useful P4.14: EORTC RANDOMIZED PHASE II STUDY OF EXTENDED in better patient stratification. There is a certainly a need for a larger PLEURECTOMY/DECORTICATION (E-PD) PRECEDED OR randomized study before denying patients of a well established surgical FOLLOWED BY CHEMOTHERAPY IN PATIENTS WITH EARLY STAGE alternative and multimodality treatment. MALIGNANT PLEURAL MESOTHELIOMA (MPM)

Disclosure: No significant relationships. Loic Lang-Lazdunski1, Jan P. Van Meerbeeck2 1Thoracic Surgery, Guy’s And St Thomas’ Nhs Foundation Trust, London/ UNITED KINGDOM, 2Thoracic Oncology, Ghent University Hospital, Gent/ BELGIUM

Background: Extra-pleural pneumonectomy (EPP) is the most commonly used surgical procedure in MPM, but a recent randomized feasibility study suggested no benefit and a possible harm (Treasure 2011). Uncontrolled series suggest that lung sparing surgical approaches such as e-PD might result in equal to better outcome (Lang-Lazdunski 2011; Flores, 2008) and

iMig2012.org • Abstract Book 135 should be further investigated. The rate of major morbidity was significantly higher in mesothelioma of the right hemithorax compared to the left side (Fisher’s Exact Test: 0.006) as Methods: Selected patients with MPM will be randomly allocated BPF was only observed on the right side. The 30 day mortality did not differ to e-PD preceded or followed by 4 cycles of cisplatin/pemetrexed at significantly in right-sided compared to left-sided EPP. The overall survival standard doses. Primary endpoint is the rate of success to complete (from diagnosis) of patients treated with chemotherapy and EPP was in the full treatment within 20 weeks after registration. Succes is defined both centres closed to 2 years. as any registered patient who has received 4 cycles of pemetrexed and chemotherapy, has undergone e-P/D, is alive at week 20 after registration Conclusion: Peri-operative morbidity and mortality after induction CTX without evidence of progression or relapse nor toxicity grade ≥ 3. To and EPP for MPM patients can be maintained at a reasonable level at high declare that an arm is feasible, at least 25 patients out of 32 registered volume centres with long-term experience. Surgical morbidity but not in each arm should have a succesfull treatment. Secondary endpoints are mortality is increased after right-sided EPP. process indicators of the quality and uniformity of e-P/D, progression free and overall survival, toxicity, safety and operative mortality and morbidity. Disclosure: No significant relationships. Tissue will be centrally collected and stored for molecular translational research.

Results: The trial is expected to run from January 2013 till December 2014 POSTER SESSION 4 September 13, 2012 16:00-17:00 with resulst being available in 2015 poster sessions | september 13 P4.16: CLINICAL AND PATHOLOGICAL FEATURES OF FIVE-YEAR Conclusion: This randomized trial will address the feasibilty of e-P/D in SURVIVORS OF MALIGNANT PLEURAL MESOTHELIOMA combination with standard (neo-)adjuvant chemotherapy and set the experimental arm of an ensuing phase III trial Taiichiro Otsuki1, Asuka Okada2, Kazuya Fukuoka2, Shingo Kanemura1, Eisuke Shibata3, Eriko Masachika2, Hitomi Kamiya2, Miki Honda2, Hisaya Disclosure: No significant relationships. Okuwa2, Risa Maeda2, Koji Mikami2, Yoshitaka Nogi2, Noriko Hirayama2, Takayuki Terada2, Shusai Yamada2, Kunihiro Tamura2, Chiharu Tabata2, Noriaki Tsubota4, Seiki Hasegawa5, Takashi Nakano6 1Division Of Respiratory Medicine Department Of Internal Medicine, POSTER SESSION 4 September 13, 2012 16:00-17:00 Hyogo College Of Medicine, Nishinomiya/JAPAN, 2Division Of Respiratory Medicine, Department Of Internal Medicine, Hyogo College Of Medicine, P4.15: PERI-OPERATIVE OUTCOME OF EXTRAPLEURAL Nishinomiya/JAPAN, 3Division Of Respiratory Medicine Department PNEUMONECTOMY AFTER CHEMOTHERAPY FOR MALIGNANT Of Internal Medicine, Hyogo College Of Medicine, Nishinomiya/ PLEURAL MESOTHELIOMA IN 189 PATIENTS FROM TWO JAPAN, 4Thoracic Oncology, Hyogo College Of Medicine, Nishinomiya/ INSTITUTIONS JAPAN, 5Thoracic Surgery, Hyogo College Of Medicine, Nishinomiya/ JAPAN, 6Respiratory Medicine, Hyogo College Of Medicine, Nishinomiya/ Isabelle Opitz1, Tetsuzo Tagawa2, Martina Friess1, Peter Kestenholz1, JAPAN Didier Schneiter1, Marc De Perrot3, Walter Weder1 1Division Of Thoracic Surgery, University Hospital Zurich, Zurich/ Background: The incidence of malignant pleural mesothelioma (MPM) in SWITZERLAND, 2Thoracic Surgery, Universtity Of Toronto, Toronto/ON/ Japan is predicted to increase over the next few decades. The prognosis of CANADA, 3Toronto General Hospital And Princess Margaret Hospital, MPM is poor, with the median survival reported to be approximately 9-17 Toronto/ON/CANADA months. A lot of prognostic factors are known for MPM, but still prediction of disease-related future remains unclear. The aim of this study was to Background: Reports in the literature suggest high morbidity and mortality for evaluate clinical and pathological features of five-year survivors of MPM. pleuropneumonectomy (EPP) after induction chemotherapy for mesothelioma patients. Therefore we analysed the peri-operative outcome of this patient Methods: We retrospectively obtained clinical data from the medical international mesothelioma interest group cohort in a bi-institutional setting. records of 467 patients who were diagnosed with MPM in Hyogo College of Medicine Hospital. Overall survival outcome was analyzed in 347 patients Methods: From 1999 to 2011, 189 MPM patients completed EPP after platinum- with MPM. Kaplan-Meier analysis was used to estimate the cumulative based induction chemotherapy at 2 institutions for thoracic surgery. The median survival probability. age of both patient cohorts with more than 80% male patients and the histological subtype (mostly epitheloid) as well as the stage of disease (predominantly pT3) Results: Overall median survival time (MST) of 347 patients with MPM was were comparable between both cohorts. Peri-operative mortality and major 15.8 months. Survival rates for one and two year were 59.5% and 30.0%, morbidity (pulmonary embolism (PE), bleeding, ARDS, empyema, bronchopleural respectively. Fourteen patients (4.0%) survived at least 5 years following fistula (BPF), chylothorax, patch failure, others) were analysed. initial treatments, including 4 remaining alive. Of the 5-year survivors, 7 were female, 6 had left-side disease, and the mean age was 61.0 years Results: Overall morbidity- and mortality rates for both institutions are (range 35-74). Eleven had epithelioid tumors and remaining 3 unclassified summarized in the following table. ones. Six had clinical stage I disease. All patients received systemic chemotherapy. Five underwent extrapleural pneumonectomy (EPP), and 2 pleurectomy/decortication (P/D). One received adjuvant radiotherapy. EPP after Ind Ctx (n) 189 Seven received multimodality treatment, while remaining 7 received 30d mortality 10 (5%) chemotherapy alone. The longest survival of 93 months was achieved in one female patient receiving induction chemotherapy followed by EPP. major morbidity 63 (33%) empyema / (+ BPF) 28 (15%) / (18 (9%)) Conclusion: The actual five-year survival was 4.0% in 347 patients with chylothorax 11 (6%) MPM. Epithelioid histologic subtype is identified as a favorable prognostic feature. Chemotherapy alone as well as surgery-based multimodality patch failure 8 (4%) treatment might be associated with long-term survival in patients with bleeding 7 (4%) MPM.

pulmonary embolism 5 (3%) Disclosure: No significant relationships. ARDS 3 (2%) esophageal perforation 1 (1%) cardiac arrest 1 (1%)

iMig2012.org • Abstract Book 136 POSTER SESSION 4 September 13, 2012 16:00-17:00 negatively associated with survival. We have assessed these factors in our series in a small scale hospital, Saitama Prefecture near Tokyo in Japan. P4.17: PHARMACOKINETIC ANALYSIS OF CISPLATIN DURING HYPERTHERMIC INTRATHORACIC CHEMOTHERAPY PERFUSION Methods: From 2005 to 2011, ten patients were diagnosed with (HITHOC) AFTER PLEURECTOMY AND DECORTICATION FOR MPM and treated in our hospital. Four of ten underwent extra-pleural pneumonectomy (EPP) and one of ten underwent tumorectomy because TREATMENT OF PLEURAL MALIGNANCIES of pleural dissemination. After resection, they received chemotherapy. Other four were treated with chemotherapy. Only one patients was Michael Ried1, Tobias Potzger1, Nico Braune1, Zsolt Zsiklavari2, Reiner impossible to treat because of poor performance status. Age, performance Neu1, Claudius Diez1, Berthold Schalke3, Hans-Stefan Hofmann1 status (ECOG), white blood cell count and C-reactive protein level of each 1Department Of Cardiothoracic Surgery, University Medical Center patients were investigated before histological confirmation. Three risk Regensburg, Regensburg/GERMANY, 2Department Of Thoracic Surgery, groups, (high, moderate and low risk group) were defined by the presence Hospital Barmherzige Brüder/GERMANY, 3Department Of Neurology, of following factors: old age ( 70< ), non–epithelioid type, poor University Of Regensburg At The District Medical Center/GERMANY performance status ( ０＜ ECOG ), high white blood cell count ( 9000 < Background: Cisplatin is a major drug for the treatment of pleural WBC ) and high C – reactive protein level ( 4 < CRP ). malignancies. Objectives of this study were to assess the pharmacokinetics Results: Eight of ten died within three years from the diagnosis. Two were and toxicity of intrapleural administered cisplatin during hyperthermic still alive but both of them suffered relapse. Risk groups were correlated poster sessions | september 13 intrathoracic chemotherapy perfusion (HITHOC) following pleurectomy/ with survival (below). decortication (P/D) in patients with malignant pleural mesothelioma or advanced thymoma with pleural spread (Masaoka stage IVa).

Methods: Pharmacokinetic analysis (ICP-MS) on 10 patients who received intrapleural cisplatin with a dosage of 100 mg/m2 (group 1: n= 5) or 150 mg/m2 (group 2: n= 5) at 42°C perfusate temperature. Simultaneous pleural perfusion fluid (perfusate) and serum samples were collected at the beginning and every 15 minutes during one hour of HITHOC. Subsequent serum samples were collected at the end of the operation, 6, 12 and 24 hours postoperative.

Results: There were no severe local or systemic chemotherapeutic related complications observed. In both groups mean cisplatin levels in the perfusate slightly decreased during the HITHOC procedure. The mean

area under the curve ratios (AUCperfusate:AUCserum) of cisplatin were nearly similar between both groups. The mean AUC of cisplatin in the perfusate was approximately 57 times (group 1) and 54 times (group 2) greater than detected in the serum during one hour of chemotherapy perfusion. The mean peak of cisplatin in the serum was reached after one hour of chemotherapy perfusion and after that continuously decreased within the Conclusion: In Japan, the West Part of Japan’s prognostic factors is first postoperative day. Finally, the area under the curve of cisplatin in the possible to be simple and easy method in MPM patients. serum did not significantly differ (p= 0.18) between both groups up to 24 hours after perfusion was started. Disclosure: No significant relationships.

Conclusion: Cytoreductive surgery (P/D) in combination with hyperthermic international mesothelioma interest group intrathoracic chemotherapy perfusion (HITHOC) with cisplatin is safe and feasible with the advantage of high local cisplatin concentrations which POSTER SESSION 4 September 13, 2012 16:00-17:00 might increase its efficacy but with respectable serum levels. This indicates a pharmacological advantage for this type of administration. But still there P4.19: NOVEL URINARY BIOMARKERS FOR THE EARLY DETECTION is no consensus about the optimal dosage of cisplatin and further studies OF KIDNEY INJURY FOLLOWING CYTOREDUCTIVE SURGERY AND are needed. INTRACAVITARY CISPLATIN LAVAGE FOR MESOTHELIOMA

Disclosure: No significant relationships. Sushrut Waikar1, K Annette Mizuguchi2, David J. Sugarbaker3, Joseph Bonventre1, Gyorgy Frendl4 1Medicine, Renal Division, Brigham And Women’s Hospital, Boston/MA/ UNITED STATES OF AMERICA, 2Anesthesiology Perioperative And Pain POSTER SESSION 4 September 13, 2012 16:00-17:00 Medicine, Brigham And Women’s Hospital, Boston/MA/UNITED STATES OF AMERICA, 3Division Of Thoracic Surgery, Brigham And Women’s Hospital, P4.18: THE WEST PART OF JAPAN PROGNOSTIC FACTORS IN Boston/MA/UNITED STATES OF AMERICA, 4Anesthesiology, Perioperative, MALIGNANT PLEURAL MASOTHELIOMA WAS MORE USEFUL And Pain Medicine, Brigham And Women’s Hospital, Boston/MA/UNITED THAN THE EUROPIAN ORGANIZATION FOR RESEARCH AND STATES OF AMERICA TREATMENT OF CANCER (EORTC) PROGNOSTIC SCORERING SYSTEMS IN A SMALL SCALE HOSPITAL IN JAPAN. Background: Acute kidney injury (AKI) is commonly seen with patients undergoing cytoreductive surgery with or without intracavitary cisplatin Mamoru Ueda, Katsuhiko Aoyama, Kohei Tagawa, Masashi Goto, Kozo lavage for pleural mesothelioma. The gold standard for the diagnosis of Asanuma, Shumei Kan AKI is the rise of serum creatinine (SCr) but its slow rise leads to delayed Thoracic Surgery, Higashisaitama National Hospital, Hasuda, Saitama/ diagnosis. Novel tubular injury biomarkers have been identified in JAPAN animal models of ischemic and nephrotoxic AKI to aid the early, accurate diagnosis. We hypothesized that those AKI biomarkers maybe also useful Background: The European Organization for Research and Treatment to diagnose AKI secondary to cisplatin-induced kidney injury and peri- of Cancer (EORTC) prognostic scoring systems has been available for operative stress. malignant pleural mesothelioma (MPM) patients. Recently prognostic factors in patients in West Part of Japan has been published. In this study, Methods: We have enrolled and measured urinary biomarkers pre- and patients of 70 years and above, non–epithelioid type, poor performance post-operatively from 116 individuals undergoing cytoreductive surgery, 73 status, high white blood cell count, high C – reactive protein level were of whom received intracavitary cisplatin lavage (225 mg/m2).

iMig2012.org • Abstract Book 137 Results: Post-operative AKI (defined as a > 50% rise in SCr) developed in 64 patients (29%), and 8 (7%) required renal replacement therapy. Peak post-operative urinary kidney injury molecule-1 (KIM-1 - a type- 1 transmembrane protein expressed at high levels in proximal tubular epithelial cells following ischemic or toxic injury) levels were 22.7 ng/ mg of creatinine in those with AKI and 7.8 ng/mg of creatinine in those without AKI. KIM-1 expression patterns were compared with that of other biomarkers (N-acetyl D glucosaminidase, neutrophil gelatinase-associated lipocalin, L-type fatty acid binding protein, interleukin 18, and vascular endothelial growth factor).

Conclusion: A unique pattern of urinary tubular injury biomarker expression after cisplatin exposure may provide improved diagnosis of AKI, enabling the prompt institution of renal protective strategies.

Disclosure: No significant relationships. poster sessions | september 13 international mesothelioma interest group

iMig2012.org • Abstract Book 138 Poster Session 4 Genomics September 13, 2012 16:00-17:00

POSTER SESSION 4 September 13, 2012 16:00-17:00 POSTER SESSION 4 September 13, 2012 16:00-17:00

P4.20: GENE EXPRESSION RATIO TESTS USING ULTRASOUND- P4.21: GENETIC RISK FACTORS FOR MALIGNANT PLEURAL GUIDED FINE NEEDLE ASPIRATION BIOPSIES IN A PROSPECTIVE MESOTHELIOMA (MPM): A GENOME-WIDE ASSOCIATION STUDY CLINICAL TRIAL IN MALIGNANT PLEURAL MESOTHELIOMA 1 2 3 Giuseppe Matullo , Simonetta Guarrera , Marta Betti , Giovanni poster sessions | september 13 Assunta De Rienzo1, Beow Y. Yeap2, David J. Sugarbaker1, Raphael Fiorito2, Daniela Ferrante4, Floriana Voglino2, Gemma Cadby5, Bueno1 Cornelia Di Gaetano2, Fabio Rosa2, Elisabetta Casalone3, Marina 1Division Of Thoracic Surgery, Brigham And Women’s Hospital And Harvard Padoan4, Mara Giordano3, Anna Aspesi3, Caterina Casadio3, Francesco Medical School, Boston/MA/UNITED STATES OF AMERICA, 2Department Of Ardissone6, Enrico Ruffini7, Pier Giacomo Betta8, Roberta Libener8, Medicine, Massachusetts General Hospital And Harvard Medical School, Roberto Guaschino9, Ezio Piccolini10, Lyle Palmer5, Monica Neri11, Dario Boston/MA/UNITED STATES OF AMERICA Mirabelli12, Donatella Ugolini13, Stefano Bonassi11, Corrado Magnani14, Irma Dianzani3 Background: Ultrasound-guided (US) fine-needle aspiration (FNA) biopsy 1Biology, Genetics And Biochemistry And Human Genetics Foundation, is a minimally invasive clinical technique that allows sampling of small University Of Torino, Torino/ITALY, 2Human Genetics Foundation, lesions. It is performed on a routine basis and has a major impact on the Human Genetics Foundation, Torino/ITALY, 3Dept.Of Health Sciences, therapeutic management of patients. US-guided FNAs are commonly used University Of Eastern Piedmont, Novara/ITALY, 4Dept.Of Translational to diagnose cancers and detect metastasis by providing specimens for Medicine, University Of Eastern Piedmont, Novara/ITALY, 5Ontario cytopathology and histopathology. We have previously described a gene Institute For Cancer Research, Toronto/CANADA, 6Dept.Clinical And expression ratio-based method able to translate comprehensive expression Biological Sciences, University Of Torino, Orbassano (torino)/ITALY, 7Dept. profiling data into simple clinical tests that are based on the expression Clinical Physiopathology, University Of Torino, Torino/ITALY, 8Pathology, levels of a relatively small number of genes. We also developed several Azienda Ospedaliera Ss.Antonio E Biagio E Cesare Arrigo, Alessandria/ diagnostic and prognostic gene expression tests. In this study, we applied ITALY, 9Transfusion Centre, Azienda Ospedaliera Ss.Antonio E Biagio for the first time the gene ratio technique to prospective enrolled patients E Cesare Arrigo, Alessandria/ITALY, 10Pneumology Unit, Ospedale undergoing US-guided FNAs and showed that it is a useful tool to perform Casale Monferrato, Casale Monferrato/ITALY, 11Unit Of Clinical And molecular diagnosis in MPM. Molecular Epidemiology, Irccs San Raffaele Pisana, Roma/ITALY, 12Unit Of Cancer Epidemiology, Cpo-Piemonte And University Of Turin, Torino/ Methods: One hundred and forty-nine US-guided FNA biopsies were ITALY, 13Department Of Oncology, Biology And Genetics, University Of obtained from a cohort of 56 patients at the time of definitive surgery prior Genova, Genova/ITALY, 14Translational Medicine, University Of Eastern incision with an average of three biopsies for patient, and used to isolate Piedmont, Novara/ITALY and quantify total RNA. Patients were enrolled in a clinical trial approved

by the IRB. RNA was amplified (Illumina TotalPrep RNA Amplification kit, Background: Asbestos is the main risk factor for malignant pleural international mesothelioma interest group Ambion) and analyzed by Real-Time PCR (RT-PCR). The samples were mesothelioma (MPM). Many different mechanisms have been hypothesized examined by the diagnostic tests MPM vs. normal pleura (NP) (UBE2T/ for its carcinogenic effect. Among them, ROS induced oxidative stress plays AGENCOURT _ 14535501; MAGED1/ADCY4; PAK4/MYH11) and MPM a crucial role, either directly or through activation of inflammatory cells. vs. lung adenocarcinoma (ADCA) (calretinin/claudin-7, VACb/TACSTD1, Only 10% of individuals exposed to high levels of asbestos develop MPM, MRCOX2/TITF1). When the gene ratio test was performed on multiple FNA which may be explained by individual susceptibility, including genetic biopsies obtained from an individual patient, the diagnosis was determined risk factors. Previous studies suggested a role of genetic polymorphisms by a majority rule. Diagnosis was considered as equivocal if conflicting in DNA-repair and oxidative-metabolism enzymes. These studies report results were obtained from two biopsy specimens from the same patient. significant OR of 1.4-3.5 for several SNPs and showed that genetic factors could indeed have a role in the pathogenesis of MPM, though much lower Results: We first evaluated the sensitivity of a sequential combination of than that entailed by asbestos. Conversely, dominant mutations in BAP1 diagnostic tests: MPM vs. NP followed by MPM vs. ADCA. The diagnostic have been reported to cause a new cancer prone syndrome, characterised test MPM vs. NP was applied to 120 FNA biopsies from 55 patients to by susceptibility to MPM, melanoma, and other types of cancer. The aim determine whether each FNA biopsy contained tumor cells. Twenty-nine of our work was the identification of genetic risk factors involved in the samples were not analyzed because of low amount of extracted RNA. development of MPM. Twenty-seven samples were called not-MPM and were excluded for further analysis. Ninety-three biopsies from 48 patients were classified as MPM Methods: We performed a genome-wide association study on 392 cases and were analyzed by the diagnostic test MPM vs. ADCA. Forty patients and 379 controls. A unique asset of our study is the complete clinical (83%) (95% CI: 70-93%) were called as MPM. FNA biopsies from 4 patients definition of asbestos exposures that is available for each patient and showed equivocal results, whereas 4 patients were diagnosed as ADCA. control. Cases and controls are from three towns in Northern Italy (Casale Next, we determined the sensitivity of test MPM vs. ADCA in 149 FNAs from Monferrato, Turin, Genoa). Casale is a small town exposed to a high 56 patients. We found that 49 of 56 patients (88%) (95% CI: 76-95%) asbestos pollution because of an asbestos cement factory, active in 1907- were correctly classified as MPM whereas 3 patients had FNAs showing 1986. Besides factory workers, also Casale dwellers are at increased risk equivocal results, and 4 patients were called ADCA. as compared with residents in the surrounding area. All subjects signed an informed consent form. Patients and controls were genotyped for more Conclusion: In this study, we demonstrated that the sequential than 370.000 SNPs and 21.000 CNVs, using the Illumina HumanCNV370- combination of the two diagnostic ratio tests has sensitivity comparable Quad.v3 BeadChip. to the MPM vs. ADCA test that has been validated in several independent cohorts of patients. In addition, we showed for the first time that the Results: Preliminary data show that several SNPs (located in 6q21, 5q35, combination of US-guided FNA and gene ratio test is a very useful and 19q13, 3q26, 4q32.1) were associated with MPM at a level of p=10-6/10-7 sensitive method to diagnose MPM. testing a per-allele model adjusted for gender, age, sex and PCA (Principal

Disclosure: No significant relationships. iMig2012.org • Abstract Book 139 Component Analysis) cluster in the exposed case-control study, and POSTER SESSION 4 September 13, 2012 16:00-17:00 adjusting further for asbestos exposure in the overall case-control study. Replication is on-going on a panel of 428 patients and 1269 controls P4.23: CELL CYCLE CHECKPOINT GENES AND HOMOLOGOUS from Australia (collaboration with Prof. L.Palmer). Functional and gene RECOMBINATIONAL REPAIR ARE OVEREXPRESSED IN expression studies are on-going on associated variants. MALIGNANT PLEURAL MESOTHELIOMA

Conclusion: The definition of genetic risk factors contributing to the Oluf D. Røe, Arnar Flatberg, Hans E. Krokan development of MPM may help to identify the pathophysiology of MPM and Department Of Cancer Research And Molecular Medicine, Norwegian help in the definition of the actual risk of individuals exposed to asbestos. University Of Science And Technology (Ntnu), Trondheim/NORWAY

Disclosure: No significant relationships. Background: Malignant pleural mesothelioma is an asbestos-related cancer with median survival of 12 months. It is highly resistant to most known DNA-damaging agents and irradiation. We previously showed that homologous recombination and other DNA repair systems were POSTER SESSION 4 September 13, 2012 16:00-17:00 overexpressed in mesothelioma. A genome-wide analysis with two different platforms was undertaken to further validate these findings. P4.22: SCREENING OF MPM PATIENTS FOR ACTIVATING SOMATIC MUTATIONS WITHIN PDGFR-BETA Methods: Genome-wide microarray analysis of mRNA from malignant poster sessions | september 13 pleural mesothelioma (n=6) and normal parietal pleural samples (n=7) 1 2 3 Ombretta Melaiu , Gabriella Fontanini , Barbara Costa , Laura using the HumanHT-12 v3 Expression BeadChip Kit (Cat# BD-103-0603) 2 4 2 3 1 Boldrini , Luciano Mutti , Elisa Sensi , Sara Bendinelli , Elisa Bracci , (Illumina, Inc., San Diego, CA) of 48 000 transcripts and Affymetrix 5 6 1 1 Marco Lucchi , Roberto E Favoni , Federica Gemignani , Stefano Landi Human Genome U133 Plus 2.0 GeneChip (Affymetrix, Santa Clara, CA, 1 2 Department Of Biology, University Of Pisa/ITALY, Division Of Pathological USA), 47 000 trancripts was performed according to the manufacturer’s 3 Anatomy, Department Of Surgery, University Of Pisa/ITALY, Department Of instructions. The experiments are registered in ArrayExpress according to Psychiatry, Neurobiology, Pharmacology And Biotechnology, University Of the MIAME guidelines. The two datasets were separately analysed with the 4 Pisa/ITALY, Lab Of Clinical Oncology, Hospital Of Vercelli, Borgosesia (vc)/ limma package in Bioconductor (www.bioconductor.org). Robust rankings 5 ITALY, Department Of Cardiac Thoracic And Vascular Surgery, University were produced by aggregating results of jackknifed Limma models using 6 Of Pisa/ITALY, Department Of Translational Oncology, National Cancer the GeneSelector package (www.bioconductor.org). A list of the 1500 top Institute/ITALY ranked genes from both platforms was chosen (this is an ordered list where a p-value is not available). Moreover, a ranked list of GO terms based on Background: Platelet-derived growth factor receptor beta (PDGFR-beta) the top 1500 gene-list was produced using Fischer’s exact test (TopGO and its natural ligand PDGF, as well as other tyrosine kinase receptors, play software, www.bioconductor.org). a fundamental role in growth, proliferation, and invasiveness of malignant pleural mesothelioma (MPM). In particular, it was shown that PDGFR-beta Results: Among the top GO terms the mitosis and replication-related genes is frequently activated in this neoplasm and activating mutations are often were ranked highest, with cell cycle checkpoint ranking number four and found within exon 12 and 18 of its gene. PDGFR-beta is a target for the DNA repair number 12. Among several overexpressed DNA repair genes, selective tyrosine-kinase inhibitor imatinib mesylate (STI571, Glivec) that is the homologous recombination genes RAD21, RAD51AP1, RAD54L, EME1 as currently in clinical trial combined with gemcitabine. Nevertheless, it was well as Fanconi anemia genes that recently also were assigned a checkpoint observed that a subset of MPM patients do not respond to the treatment. function. Moreover, among the checkpoint genes, TAOK1, a spindle The causes of this resistance are, at least in part, ascertained in other types checkpoint and platelet/megakaryocyte gene was highly overexpressed. of tumours. In the gastro-intestinal stromal tumours specific mutations, called “gatekeeper mutations” within exon 14 of PDGFR-beta, make cells Conclusion: DNA damage response including checkpoint and DNA repair insensitive to imatinib. To date, the frequency of activating mutations systems are highly overexpressed and overrepresented in malignant pleural within PDGFR-beta at the presentation in MPM is unknown. mesothelioma. This has been associated with treatment resistance against international mesothelioma interest group DNA damaging agents and manipulation of these systems may be an Methods: We performed a mutation screening of 100 surgically resected important path to improve the effect of chemotherapy and radiation. MPMs, to ascertain the somatic mutation frequency of PDGFRB at diagnosis. The mutation screening of the whole exons 12 and 18 of PDGFR-beta was Disclosure: No significant relationships. performed with automatic sequencing (Sanger reaction).

Results: We did not observe any mutation in all samples. POSTER SESSION 4 September 13, 2012 16:00-17:00 Conclusion: Because PDGFR-beta gene was found over-expressed at mRNA level in several cell lines and tissue specimens, we gather that MPM is P4.24: THE EXPRESSION OF LONG NONCODING RNAS IN not driven by somatic mutations but, rather, that PDGFR-beta is involved MALIGNANT PLEURAL MESOTHELIOMA in MPM because of its increased expression (that could be functionally equivalent to a constitutive activation). Then, we induced a long-term Casey M. Wright1, Michaela B. Kirschner1, Yeun Yee Cheng1, Vandana resistance to imatinib in the over-expressing PDGFR-beta human MPM Relan2, Nico Van Zandwijk1, Glen Reid1 established cell line MERO-14. Once resistant clones are obtained, they will 1Asbestos Diseases Research Institute, University Of Sydney, Concord/ be screened for mutations in exons 12, 14, and 18. The analysis of resistant AUSTRALIA, 2Department Of Thoracic Medicine, The Prince Charles clones could help in revealing whether PDGFR-beta plays a role in the Hospital, Brisbane/QLD/AUSTRALIA secondary resistance to imatinib. Background: Long noncoding RNAs (lncRNA) are a class of RNAs >200 Disclosure: No significant relationships. nucleotides in length, that do not code for protein but make up >90% of the human genome. Recent studies have investigated the potential role of lncRNAs in cancer. For example, HOTAIR and MALAT1 are implicated in metastases of lung and breast cancer. As malignant pleural mesothelioma (MPM) is an aggressive disease with poor prognosis and lncRNAs have not been previously investigated, our aims were to characterize the expression of lncRNAs potentially involved in MPM biology.

Methods: To identify novel lncRNAs involved in MPM, microarray profiling was performed on five cell lines - the immortalized normal mesothelial

iMig2012.org • Abstract Book 140 cell line (MeT-5A) and four MPM lines (two epithelioid H28 and H226 and two biphasic MM05 and MSTO) using Invitrogen’s NCode long noncoding RNA microarrays which allow simultaneous assessment of mRNAs and lncRNAs. Data was feature extracted using proprietary software and preprocessed using Gene Spring V12.0. High priority candidate lncRNAs were selected on the basis of statistical (P<0.05) and biological significance (>3-fold difference). Expression levels of candidate mRNA and lncRNAs were validated using Taqman Gene Expression assays on 7 MPM cell lines and 5 cancer cell lines (HCT115, HCT116, A549, PC-3, MDA-MB-231, MCF-7).

Results: Microarray profiling of MeT-5A versus MPM cell lines identified 350 probes (310 mRNA, 40 lncRNA) differentially expressed between mesothelioma and normal cell lines at >3-fold and P<0.05. These probes included known cancer genes including EGFR, CDKN2A, MYC andMET, all of have been associated with MPM. In addition we identified PLAUR, a gene previously shown to be highly expressed in human mesothelial cells. The majority of candidates were found to be up-regulated in tumor cell lines. Stratification by histological subtype identified 172 probes differentially poster sessions | september 13 expressed between biphasic and epithelioid cell lines. Interestingly, 46 probes were found to overlap between normal versus cancer and epithelioid versus biphasic analyses. Validation of microarray data by RT-qPCR is underway and will be presented at the conference. Finally bioinformatics analyses identified 40 lncRNAs with a putative role in MPM, including NEAT1 which has been implicated in ovarian cancer. RT-qPCR analysis also demonstrated increased expression of MALAT1 in MPM cell lines compared to MeT-5A. The most pronounced changes were observed in the three epithelioid lines H2052 (fold change 43.9), H2452 (fold change 8.4) and H226 (fold change 4.4). For the remaining cell lines (REN, H28, MSTO, MM05) relatively small differences were observed (<2-fold change).

Conclusion: Microarray profiling has identified novel lncRNAs and mRNAs with a putative role in MPM biology. Independent validation of MALAT1 using RT-qPCR identified over-expression in MPM cell lines compared to a normal mesothelial cell line MeT-5A. Further biological and functional validation is required to confirm the role of novel lncRNAs in the biology of MPM.

Disclosure: No significant relationships. international mesothelioma interest group

iMig2012.org • Abstract Book 141 Poster Session 4 Chemotherapy and Drug Selection September 13, 2012 16:00-17:00

POSTER SESSION 4 September 13, 2012 16:00-17:00 Methods: The clinical records of elderly pts (≥70 years old) with MPM referred from January 2005 to November 2011 were reviewed. For each P4.25: ABCB1 IS A CRITICAL REGULATOR OF VINORELBINE patient, age and gender, histology, Eastern Cooperative Oncology Group INDUCED APOPTOSIS Performance Status (ECOG-PS), Charlson Comorbidity Index (CCI) and treatment modalities were collected. The study endpoint was overall 1 2 3 Sara Busacca , Jaine Blaney , Dean Fennell survival (OS). poster sessions | september 13 1University Of Leicester, 9hn/UNITED KINGDOM, 2Centre For Cancer Reasearch And Cell Biology, Queen’s University Of Belfast, 7bl/UNITED Results: Out of a total of 610 cases, 210 elderly pts were identified KINGDOM, 3University Of Leicester, Hn/UNITED KINGDOM (34% of the whole MPM population observed in the study period). Pts characteristics were: median age 75 yrs (range 70-92), M/F 132/78, Background: Vinorelbine has been shown to exhibit useful clinical activity epithelial/non-epithelial histology 140/70, ECOG-PS 0/1/2/unknown in malignant pleural mesothelioma (MPM). A UK randomized clinical trial 130/67/9/4. CCI was 0 in 128 pts (61%), ≥4 in 59 pts (28%). Treatment was is planned to evaluate vinorelbine in mesothemioma (VIM). Identification multimodality therapy including surgery in 16, chemotherapy in 153 (73%) of predictive biomarkers may be essential for successful development of and best supportive care only in 41 pts (19%). Chemotherapy was mainly vinorelbine in mesothelioma. pemetrexed-based. Median OS was 11.0 months. In a multivariate model, non-epithelial histology, age ≥75 yrs and the presence of co-morbidities Methods: We selected two cell lines with resistance to vinorelbine, according to CCI (HR 1.15; 95% CI 1.07-1.23, p<0.001) were all significantly followed by gene expression microarray analysis. Functional genetics and correlated to a shorter OS. In the same model, treatment with pemetrexed pharmacological inhibitor studies were conducted to validate putative was associated with improved OS. Age and co-morbidity were not biomarker ABCB1 as a regulator of vinorelbine sensitivity. significantly correlated.

Results: Gene expression analysis revealed ABCB1 as one of the mostly Conclusion: Pemetrexed-based chemotherapy is feasible in selected highly overexpressed genes in the resistant cells as confirmed by western elderly pts with MPM. Comorbidity is a significant prognostic factor, and blot. We then utilized inhibitors of ABCB1 to evaluate the effect on should be carefully considered in patient selection. Prospective dedicated vinorelbine induced apoptosis. XR9051 or verapamil when combined with trials in elderly pts with MPM selected according to comorbidity scales are vinorelbine markedly increased PARP and caspase 9 activation, but were warranted. inert as single agents in the resistant MPM cells. Resistance to vinorelbine was also reversed by RNAi silencing of ABCB1. Disclosure: No significant relationships.

Conclusion: MPM cells evolve upregulation of ABCB1 during selection for vinorelbine resistance. Significant focal amplification of ABCB1 is reported POSTER SESSION 4 September 13, 2012 16:00-17:00 in solid tumours however the frequency in mesothelioma is being explored. international mesothelioma interest group Our data suggest that ABCB1 is a putative predictive biomarker warranting evaluation in the VIM trial. P4.27: HIGH INCIDENCE OF HYPER-SENSITIVITY REACTIONS IN PATIENTS WITH MALIGNANT PLEURAL MESOTHELIOMA RE- Disclosure: No significant relationships. TREATED WITH CARBOPLATIN/PEMETREXED

Sergio Stinco, Cristina Ripa, Chiara M. La Spina, Maria G. Sauta, Michela Squadroni, Piermario Salvini, Giordano D. Beretta, Giovanni L. Ceresoli POSTER SESSION 4 September 13, 2012 16:00-17:00 Oncology, Humanitas Gavazzeni Clinic, Bergamo/ITALY

P4.26: MALIGNANT PLEURAL MESOTELIOMA (MPM) IN ELDERLY Background: In a prior study (Ceresoli, Lung Cancer 2011) we have PATIENTS: RESULTS OF A MULTICENTER SURVEY. proposed re-treatment with pemetrexed-based chemotherapy (PBC), mainly the combination of pemetrexed/carboplatin, as an option in Giovanni L. Ceresoli1, Federica Grosso2, Paolo A. Zucali3, Giulia Pasello4, patients (pts) with malignant pleural mesothelioma (MPM) achieving Daniela Degiovanni5, Fabio De Vincenzo3, Sergio Stinco1, Valentina a durable disease control with first-line PBC. Studies in gynecologic Polo4, Maria G. Sauta1, Carlo Dipietrantonj6, Laura Giordano3, Adolfo malignancies have shown that repeated treatment with carboplatin Favaretto4, Armando Santoro3, Mario Botta7 increases the incidence of hypersensitivity reactions (HSRs). In this 1Oncology, Humanitas Gavazzeni Clinic, Bergamo/ITALY, 2Oncologia, retrospective study we have analyzed the occurrence of HSRs to Ospedale Civile, Alessandria/ITALY, 3Oncology, Humanitas Cancer carboplatin in a subset of re-treated MPM pts. Center, Rozzano, Milan/ITALY, 4Medical Oncology, Istituto Oncologico Veneto, Padua/ITALY, 5Palliative Care Unit, S. Spirito Hospital, Casale Methods: The clinical records of patients retreated with pemetrexed/ Monferrato/ITALY, 6Servizio Epidemiologia, Asl Alessandria, Alessandria/ carboplatin as second-line or further-line therapy referred to our Institution ITALY, 7Oncology, S. Spirito Hospital, Casale Monferrato/ITALY were evaluated. HSRs to carboplatin and their management were reported.

Background: The incidence of malignant pleural mesothelioma (MPM) in Results: Between October 2006 and December 2011, 18 patients (11 elderly patients (pts) is increasing in Western Countries. Elderly pts with males and 7 females) received re-treatment with pemetrexed/carboplatin. MPM are under-represented in clinical trials, and there are no specific Median age was 66 years (range 37-78). Disease control rate in this highly guidelines for their management. The aim of this study was to perform selected population was 78%. Median progression-free survival after re- a retrospective survey on this patient population in four Oncology treatment was 8.2 months (range 1.3-20.4). Seven pts received a further Departments with high MPM accrual and expertise. PBC as third-line treatment. Overall, 6 pts (33%) experienced an HSR to carboplatin after a median of 9 cycles (range 8-13) and of 18.5 months

iMig2012.org • Abstract Book 142 (range 13-45) after first carboplatin administration. All HSRs were classified apoptosis in chemoresistant cells and prevent tumor growth in mouse as grade 2 and were easily managed with steroids and anti-histaminics. models. Carboplatin administration was omitted in subsequent cycles. Disclosure: No significant relationships. Conclusion: Re-treatment with PBC in selected MPM pts is a valuable strategy, however clinicians should be aware of the high incidence of HSRs to carboplatin in this setting. Premedication and desensitization strategies should be implemented. POSTER SESSION 4 September 13, 2012 16:00-17:00

Disclosure: No significant relationships. P4.29: VINORELBINE IN MESOTHELIOMA (VIM): A RANDOMISED PHASE II TRIAL OF ORAL VINORELBINE AS SECOND-LINE THERAPY FOR PATIENTS WITH MALIGNANT PLEURAL MESOTHELIOMA (MPM) EXPRESSING BRCA1 – A STUDY IN POSTER SESSION 4 September 13, 2012 16:00-17:00 PROGRESS P4.28: THE ROLE OF TGF-ALPHA IN THE RESISTANCE TO Dean Fennell1, Angela Casbard2, Lisette Nixon3, Sara Busacca1, Jason MESOTHELIOMA CHEMOTHERAPY Lester3, Gareth Griffiths3 poster sessions | september 13 1University Of Leicester, Leicester/UNITED KINGDOM, 2Velindre Hospital/ Chrisostome Costa1, Fabian Vandermeers2, Sathya Neelature UNITED KINGDOM, 3Wales Cancer Clinical Trials Unit/UNITED KINGDOM Sriramareddy2, Céline Mascaux3, Arnaud Scherpereel4, Philippe Delvenne5, Pascale Hubert6, Luc Willems7 1 Background: Mesothelioma is increasing worldwide. However there is Cellular And Molecular Biology, University Of Liège (Ulg)-Gembloux no approved therapy in the second-line setting. Vinorelbine exhibits Agro-Bio Tech, Gembloux/BELGIUM, 2Molecular And Cellular Epigenetics, 3 promising activity, however there has been no randomised evaluation or Giga-University Of Liège (Ulg)/BELGIUM, Thoracic Oncology, Bordet validation of biomarkers to support patient stratification. We have recently Institute, Ulb/BELGIUM, 4Pneumology And Thoracic Oncology, Lille 5 reported that BRCA-1 is an essential regulator of mesothelioma sensitivity Hospital/FRANCE, Experimental Pathology, Giga-University Of Liège, to vinorelbine, and its expression is lost in approximately 38%1. The Cancer Liège/BELGIUM, 6Experimental Pathology, Giga-University Of Liège/ 7 Research UK VIM trial is to be sponsored by the University of Leicester in BELGIUM, Molecular And Cellular Epigenetics (Giga) And Molecular Biology collaboration with the Wales Cancer Clinical Trials Unit. To evaluate the (Gxabt), Giga-University Of Liège (Ulg)-Gembloux Agro-Bio Tech/BELGIUM efficacy of second-line vinorelbine plus active symptom control (ASC), versus ASC. Secondary endpoints: tolerability, response rate, change in Background: The impact of chemotherapy on the outcome of patients tumour volume and overall survival. BRCA1 expression IHC will be evaluated with malignant pleural mesothelioma (MPM) is still controversial. The as a stratification factor. median survival post diagnosis is invariably about 8 to 12 months. We hypothesized that unresponsiveness to chemotherapy is due to inadequate Methods: An open label, randomised trial of weekly vinorelbine 80mg/ gene expression in tumor cells. We have previously shown that inhibitors m2 plus ASC versus ASC alone. The control arm of ASC will be defined of histone deacetylases (such as valproate, VPA) significantly increases the by local practice. Both study arms will be continued until documented efficacy of compounds used in chemotherapy (Vandermeers et al, 2010, evidence of radiological progression or unacceptable toxicity. Clinical Cancer Research 15: 2818). A recent clinical trial on 45 relapsing MPM patients has shown that VPA in combination with doxorubicin Results: The sample size has been calculated using the parameters; increases response rates and improves quality of life in 25% patients with α=0.2, β=0.1 (90% power), hazard ratio 0.65, 1-sided logrank test and MPM (Scherpereel et al, 2011, European Respiratory Journal 37:129). 2:1 randomisation favouring vinorelbine. This requires recruitment of 114 patients. However, as we hypothesis that BRCA-1 expression is required for Methods: Using Agilent microarrays, we compared the transcriptome of vinorelbine activity and have estimated its absence in one third of patients, international mesothelioma interest group two types of cell lines (M14K and H28). Bioinformatic analyzes (Ingenuity) the sample size may be inflated to 171 patients depending on the results of identified the most relevant candidate genes. Expression of transforming an interim analysis. growth factor-alpha (TGFa) was validated by RT-qPCR and ELISA. TGFa expression level was modulated negatively by RNA interference and Conclusion: We aim to open the study in Q1 2013 and recruit over 18 positively by transfection of a cDNA vector. TGFa signaling was inhibited months. The results of this study will be used to inform the design of a with EGFR tyrosine kinase inhibitors (gefitinib and erlotinib). Apoptosis future phase III study, with stratification of patients to optimise efficacy. was assessed by different techniques (DNA fragmentation, Annexin V externalization and caspase activity). TGFa-EGFR signaling was Disclosure: No significant relationships. characterized by western blot using antiphosphopeptide antibodies. Combination chemotherapy was investigated in two mouse models (ZL34 in SCID and AB1 in Balb/c). POSTER SESSION 4 September 13, 2012 16:00-17:00 Results: To study the mechanisms associated with response to chemotherapy, we compared two types of cell lines (M14K and H28) P4.30: TREATMENT AND SURVIVAL ANALYSES OF MALIGNANT characterized by a difference in sensitivity to doxorubicin + VPA. We MESOTHELIOMA IN JAPAN observed that the basal expression level of TGFα was higher in “resistant” H28 compared to “sensitive” M14K cells. To evaluate the functional Nobukazu Fujimoto1, Kenichi Gemba1, Keisuke Aoe2, Katsuya Kato3, relevance of TGFα, we modulated its expression either by RNA interference Yukio Takeshima4, Kouki Inai4, Takumi Kishimoto1 or by transfection of a cDNA vector. Our data shows that a decrease of 1Respiratory Medicine, Okayama Rosai Hospital, Okayama/JAPAN, 2Medical TGFα expression correlated with induction of apoptosis. Inversely, an Oncology, Yamaguchi-Ube Medical Center, Ube/JAPAN, 3Radiology, inhibition of apoptosis occurred when TGFα was over-expressed. Since Okayama University Hospital, Okayama/JAPAN, 4Pathology, Nstitute Of TGFa is the ligand of EGFR, we tested the effect of gefitinib and erlotinib Biomedical & Health Sciences Hiroshima University, Hiroshima/JAPAN in combination with VPA+doxorubicin. Both EGFR inhibitors increased VPA+doxorubicin apoptosis in H28 chemoresistant cells. Finally, the new Background: There are few reports concerning treatment strategies and combination therapy VPA+doxorubicin+erlotinib prevented tumor growth their contributions to survival of patients with malignant mesothelioma in mice. (MM) in Japan.

Conclusion: Our data demonstrates that TGFα is involved in Methods: We extracted all death cases due to MM between 2003 and chemoresistance to VPA+doxorubicin, a second-line regimen for MPM. 2008. The diagnosis of MM was confirmed in 929 cases including 396 Although inefficient alone, tyrosine kinase inhibitors synergize to induce (55.9%) epithelioid type, 154 (21.7%) sarcomatoid type, 126 (17.8%)

iMig2012.org • Abstract Book 143 biphasic, and 33 (4.7%) others.

Results: Median overall survival (OS) of all MM cases was 7.7 months (95% confidence interval, 7.1-8.3). Median OS of patients with epithelioid MM was significantly longer than that of patients with biphasic (P=0.030) or sarcomatoid (p<0.001) MM. Surgical resection was performed in 172 patients (18.5%) and 449 (48.3%) received systemic chemotherapy. Survival of patients treated with both surgery and systemic chemotherapy was favorable. Median OS of patients in the late phase of the study period (2006-2008) was significantly longer than that in the early phase (2003- 2005) (8.1 vs. 7.5 months, p=0.008). Age younger than 70 years, female gender, epithelioid subtype, and clinical stage I-III were independent favorable prognostic factors. Multivariate analysis confirmed that radical surgery and systemic chemotherapy contribute to longer survival of patients with pleural MM.

Conclusion: The prognosis of MM is poor in Japan, though survival tends to prolong. poster sessions | september 13

Disclosure: No significant relationships.

POSTER SESSION 4 September 13, 2012 16:00-17:00

P4.31: THIRD LINE CHEMOTHERAPY IN ADVANCED MALIGNANT Conclusion: Sarcomatoid subtype patients had poor survival from start PLEURAL MESOTHELIOMA (MPM): SUPERIOR OUTCOME FOR of 3rd line and should probably not receive this treatment. Elderly patients FEMALES aged 70 years or above had similar prognosis as younger patients and should not generally be excluded from treatment. Female patients had Jens Benn Sorensen, Jan Nyrop Jakobsen superior prognosis. More active treatment options for relapsed MPM Dept. Oncology, Finsen Centre/National University Hospital, Copenhagen/ patients are sorely needed. DENMARK Disclosure: No significant relationships. Background: While standard treatment in 1st line is platinum based doublet chemotherapy there are no established 2nd or 3rd line treatment in advanced MPM. Data on outcome from 3rd line treatment are exceedingly rarely reported. Our group have previously published high activity in 1st line POSTER SESSION 4 September 13, 2012 16:00-17:00 treatment with carboplatin + gemcitabine + liposomized doxorubicin (J Thorac Oncol 3: 1325-31, 2008) and this regimen (CCG regimen) was hence P4.32: CHARACTERIZATION AND DRUG SENSITIVITY OF rd explored in 3 line. MALIGNANT MESOTHELIOMA CELLS FROM PLEURAL EFFUSIONS

st Methods: All patients had received 1 line treatment with either platinum Adam Szulkin1, Rita Ötvös2, Anders Hjerpe1, Laszlo Szekely2, Katalin nd + vinorelbine or platinum + pemetrexed, while 2 line was either Dobra1 international mesothelioma interest group pemetrexed or vinorelbine monotherapy. Patients had advanced MPM, 1Department Of Laboratory Medicine, Karolinska Institutet, Stockholm/ performance status 0-2, and normal organ renal, hepatic, cardiac, and SWEDEN, 2Department Of Microbiology Tumor And Cell Biology, Karolinska hematological function. Treatment was carboplatin AUC5 and liposomized Institutet, Stockholm/SWEDEN doxorubicin 30 mg/m2 i.v. day 1 q 3 wks, while gemcitabine 1000 mg/m2 was administered i.v. days 1 and 8 q 3 wks. Background: Patients with malignant mesothelioma have a poor prognosis and less than 50% respond to standard treatment (Pemetrexed and Results: Totally 53 patients were treated from June 2006 through May Cisplatin). Furthermore, patients responding to this treatment only have st 2012, representing 20.2% of patients who started 1 line treatment in an increased survival of a few months. We hypothesize that the drug the same period. Forty-seven patients were males and 6 (11%) were resistance pattern is individual for each malignant mesothelioma patient females. Median age was 66 years (range 37-76 years). Epitheloid subtype and correlates to the in vitro drug sensitivity of primary tumour cells. occurred in 70%, 6% and 24% had sarcomatoid and biphasic subtypes, The clinical outcome of these patients can then be predicted by this respectively. Forty-seven percent received 3 or more treatment courses, measurement of drug sensitivity and complemented by evaluating the maximum was 6 courses received by 8% of patients. Overall survival expression of ERCC1 and RRM1, two proteins involved in chemoresistance. from initial histologic diagnosis was median 23.7 months (range 7.6-59.9 months), while 1-, 2-, and 3-years survival rates from diagnosis were 74%, Methods: Pleural effusions containing primary malignant mesothelioma rd 28%, and 13%, respectively. Median overall survival from start of 3 line cells were received from the diagnostic routine. Cells were seeded in treatment was 7.6 months (range 0.6-41.9 months) and 1-year survival rate a 384-well plate for a robotized ex vivo testing of drug sensitivity. In rd 23%. Survival from start of 3 line was median 10.2 months for females total, 30 different drugs were tested (2 topoisomerase inhibitors, 5 (range 1.9-41.9) and 6.8 months for males (range 0.6-18.9) (p=0.026, alkylating agents, 5 antimicrotubule agents, 1 proteasome inhibitor, 10 figure 1). Median survivals were 8.5 months for epiteloid subtype (range antimetabolites, 6 antitumor antibiotics and 1 corticosteroid). Each drug 0.6-41.9), 4.7 for sarcomatoid (1.1-5.2), and 6.0 for biphasic (1.1-13.57) was tested at concentrations covering the clinically relevant span. The (p=0.399). Patients younger and elder than 70 years had similar prognosis primary cells were also further characterized by immunocytochemistry and (p=0.237). Response data will be updated at the meeting. qRT-PCR to evaluate the proportion of malignant cells in each sample and to study the expression of RRM1 and ERCC1.

Results: So far we have analyzed, characterized and tested 18 samples from 13 different patients. The samples consist of 10-100 % malignant cells and have a variable expression of ERCC1 and RRM1. Among the tested established drugs, Actinomycin-D was the most effective, with cytotoxic effects in a majority of the 18 primary cell cultures. Among the different

iMig2012.org • Abstract Book 144 groups, the antimicrotubule agents (Docetaxel, Paclitaxel, Vinblastine, POSTER SESSION 4 September 13, 2012 16:00-17:00 Vincristine and Vinorelbine) seemed to affect most samples. The sensitivity patterns varied greatly between the different primary cell cultures. Five of P4.34: VARIABILTY OF BLOOD AND CYTOKINE MARKERS the samples were resistant to most of the tested drugs and even the most IN NORMAL AND DRUG RESISTANT IL-45 CELLS IN A RAT sensitive cells were resistant to more than 50 % of the drugs. The primary MESOTHELIOMA MODEL. cell samples that were more sensitive were received from patients with a longer survival time. Chris Weir1, Amanda Hudson2, Lydnsey Peters3, Nick Pavlakis4, Stephen Clarke1 Conclusion: Our experimental approach allows us to characterize primary 1Medical Oncology, Bill Walsh Cancer Research Labs/Kolling Institute, malignant mesothelioma cells, evaluate their expression of central Sydney/NSW/AUSTRALIA, 2Medical Oncology, Bill Walsh Cancer Research drug resistance proteins and the robotized assay allows a simultaneous Labs/Kolling Institute, NSW/AUSTRALIA, 3Kolling Institute, Northern Blood determination of chemosensitivity to 30 different drugs. The obtained Research Centre, NSW/AUSTRALIA, 4Medical Oncology: Royal North Shore drug sensitivity patterns and protein levels of ERCC1 and RRM1 vary greatly Hospital, Bill Walsh Cancer Research Laboratories, Sydney/NSW/AUSTRALIA between different malignant mesothelioma samples, motivating the further development of this technique for clinical use. Background: Malignant mesothelioma (MM) is a rare and aggressive tumour with poor response rates and no curative treatment available Disclosure: No significant relationships. with most patients dying within 10-17 months of their first symptoms. The majority of patients with MM are diagnosed in stage III/IV of the poster sessions | september 13 disease when administration of systemic chemotherapy represents the only treatment option apart from palliative care. However due to the POSTER SESSION 4 September 13, 2012 16:00-17:00 inherent resistance of this disease, be it intrinsic or acquired relatively poor response to treatment is seen and relapse rates remain high. P4.33: NOVEL HDAC/DNMT TWIN INHIBITOR AS ANTICANCER Drug resistance in MM has not been extensively studied even though AGENT FOR MALIGNANT PLEURAL MESOTHELIOMA chemotherapy resistance is widely acknowledged. Blood markers such as elevated neutrophil to lymphocyte ratio (NLR) have been shown to predict Fabian Vandermeers1, Pascale Hubert2, Sathya Neelature worse prognosis but other cell types have not been tested. We developed Sriramareddy1, Julie Braun3, Irving Boittiaux4, Eric Stern4, Julie Horion2, a multicolour flow cytometric assay to look at 7 cell types (including NLR) Philippe Delvenne2, Didier Lambert4, Johan Wouters3, Luc Willems1 from a small 25µl sample of blood. These cell types include T4, T8, natural 1Molecular And Cellular Epigenetics, University Of Liège, Liège/ killer (NK) cells, B cells, neutrophils, lymphocytes and monocytes. The BELGIUM, 2Experimental Pathology, University Of Liège, Liège/ aim of these experiments was to monitor the level of these cells during BELGIUM, 3Biological Chemistry, University Of Namur/BELGIUM, 4University pleural tumour progression in normal and drug resistant IL-45 cells in our Of Louvain/BELGIUM rat mesothelioma model. Plasma cytokine profiles were also compared between groups. Background: Standard chemotherapeutic regimens are marginally efficient in malignant pleural mesothelioma (MPM) because tumor cells Methods: To look for blood cell and cytokine markers of resistance, are particularly resistant to radiotherapy and/or chemotherapy. Previous Fischer F344 rats were injected with 0.5x106 normal or drug resistant evidence indicates that unresponsiveness of tumors to conventional IL-45 rat mesothelioma cells in the pleural cavity. Drug resistant cell lines therapeutic agents might be due to inappropriate epigenetic modifications. include Cisplatin, Pemetrexed, Gemcitabine , Vinorelbine and combination Consistently, the HDAC inhibitor valproic acid (VPA) partly ameliorates the (Cisplatin/pemetrexed). Whole blood was collected twice weekly via tail efficacy of the first and second line treatments of MPM. In this perspective, bleeds and monitored for NK, T4, T8, B cell, lymphocyte, neutrophil and we aimed at identifying improved compounds affecting different epigenetic monocyte levels during disease progression by flow cytometry. Differences processes such as DNA methylation and histone deacetylation. in plasma cytokine profiles between normal IL-45 and drug resistant IL-45 cells were also screened at endpoint. Methods: Docking simulations were performed with a series of compounds international mesothelioma interest group and DNA methyltransferases (DNMTs) or histone deacetylases (HDACs). Results: NLR was only significantly higher in rats with normal IL-45 cells A lead compound called ES8 was designed and synthesized. The and combination resistant cells. NLR was also not predictive for tumour antimetabolic and proapoptotic activity of ES8 was tested in a series of volume or other factors such as rapid health deterioration such as weight MPM cell lines. The anticancer potential was tested in 2 mouse models. loss. A decrease in B cell numbers was significant in all groups except rats with combination resistant cells and proved the most consistent marker of Results: We have designed, synthesized and tested the anticancer disease progression. General trends showed a decrease in T4 and T8 cells potential of a novel compound called ES8 having concomitantly intrinsic as disease progressed, however T4/T8 ratios did not decrease in all drug HDAC and DNMT inhibitory activities. Docking simulation analyzes show resistant groups. Rapidly increasing monocyte levels were prognostic for that ES8 can interact with HDAC and DNMT catalytic pockets. ES8 is pro- rapid tumour growth and weight loss apoptotic in a panel of MPM cell lines and prevents tumor growth in mouse models. ES8 treatment is associated with histone hyperacetylation and Conclusion: We have developed a useful blood screening test for reduced DNA methylation in tumors. monitoring cells from whole blood in rats which could easily be adapted to human patients. Standard NLR monitoring maybe useful in non drug Conclusion: Our data thus shows that a dual HDAC/DNMT inhibitor may be resistant IL-45 cells but was not elevated significantly in rats with 3 out of useful to improve treatment of MPM. 4 drug resistant cell lines. This suggests that with drug resistant disease blood cell markers such as NK or B cells may be useful prognostically when Disclosure: No significant relationships. standard NLR fails. Further work into using this blood test while rats with mesothelioma are treated with chemotherapy are currently in the pipeline.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 145 Poster Session 5 Cancer Biology, Gene Regulation and Pathways and Apoptosis September 14, 2012 11:30-12:30

POSTER SESSION 5 September 14, 2012 11:30-12:30 POSTER SESSION 5 September 14, 2012 11:30-12:30

P5.01: OVERCOMING RESISTANCE TO EGFR PATHWAY INHIBITION P5.03: MICRORNA-223 TARGETS STATHMIN AND REGULATES IN MESOTHELIOMA CELLS CELL MIGRATION IN MALIGNANT MESOTHELIOMA

1 2 2 3 Vijay Agarwal , Brendan O’Sullivan , Phillipe Taniere , Michael J. Lind , Kimberly A. Birnie, Philip J. Thompson, Steven E. Mutsaers, Bahareh poster sessions | september 14 Lynn Cawkwell3 Badrian 1Department Of Cancer Studies, Hull York Medical School, Hull/ Lung Institute Of W.A. University Of Western Australia, Perth/WA/ UNITED KINGDOM, 2Queen Elizabeth Hospital, Birmingham/UNITED AUSTRALIA KINGDOM, 3Postgraduate Medical Institute, University Of Hull, Hull/UNITED KINGDOM Background: Malignant Mesothelioma (MM) is an aggressive cancer associated with asbestos exposure. MM has a poor prognosis and current Background: EGFR, MTOR and COX2 are upregulated in malignant pleural therapies and treatments remain ineffective. Australia has one of the mesothelioma (MPM) and are potential targets for directed therapy. In world’s highest rates of MM, therefore it is essential to develop a more this study we aimed to determine the cytotoxic effect of EGFR, MTOR and comprehensive understanding of this disease. Recent studies have COX2 inhibitors. identified microRNAs (miRNAs), a family of small, single stranded RNAs which regulate gene expression, as important in cancer pathogenesis. Methods: The COX2 positive cell lines MSTO-211H, NCI-H2052, In preliminary studies we identified miRNA-223 (miR-223) as aberrantly NCI-H2452 (mesothelioma) and A549 (lung cancer) were utilised. The EGFR expressed in MM. miR-223 has been associated with the pathogenesis of and PTEN status of the cell lines was determined using flow cytometry other cancers and targets the microtubule regulator stathmin, which has and immunoblotting respectively. All cell lines were tested for EGFR, KRAS been linked to cancer cell migration. Our aim was therefore to test the and BRAF mutations. Cells were incubated with single agent Cetuximab, hypothesis that miR-223 is functionally significant in MM. Gefitinib, Rapamycin, Ku0063794 (MTOR kinase inhibitor) and Celecoxib for 72 hrs and analysed using the MTS assay. Subsequently Cetuximab Methods: The expression of miR-223 and stathmin mRNA was determined and Gefitinib were combined in turn with Rapamycin, Ku0063794 and using real- time PCR in human and mouse MM cell lines and control Celecoxib. The concentration of Cetuximab (1.6 μM) and Gefitinib (1.4 mesothelial cells. stathmin and protein levels were measured by western μM) was calculated based on the steady state plasma concentrations at blot. Cells were transfected with the miR-223 precursor construct to FDA approved dosing. In each experiment 6 replicates were used per drug modulate the levels of miR-223 and the impact on stathmin expression, cell concentration and the experiment was repeated at least twice. proliferation and migration examined.

Results: All cell lines demonstrated >98% positivity for EGFR. PTEN was Results: miR-223 was expressed at significantly lower levels (p<0.01) in

absent in the MSTO-211H cells. A549 cells had a KRAS missense mutation at the MM cells compared to control mesothelial cells. In the same cell lines, international mesothelioma interest group codon 12. No other EGFR, KRAS and BRAF mutations were identified in any stathmin mRNA and protein were significantly higher (p<0.05) with a strong of the cell lines. Cetuximab showed 50% cell growth inhibition in MSTO- inverse correlation between the levels of miR-223 and stathmin. Over- 211H cells at a concentration of 1.6 µM. All other cell lines were resistant to expressing miR-223 reduced stathmin mRNA and protein, confirming that Cetuximab. All cell lines were resistant to Gefitinib at concentration <1.4 miR-223 targets and regulates stathmin in MM. Over-expressing miR-223 μM. Rapamycin and Ku0063794 demonstrated 50% cell growth inhibition did not affect cell proliferation but did cause a decrease in cell migration. in NCI-H2052, NCI-H2452 and A549 cells. Celecoxib demonstrated 50% cell growth inhibition in all cell lines. Cetuximab and Gefitinib were Conclusion: miR-223 targets stathmin and is important in regulating MM combined in turn with Rapamycin, Ku0063794 and Celecoxib. Cetuximab cell migration in vitro. when combined with Celecoxib (NCI-H2052, NCI-H2452 and A549 cells) and Ku0063794 (MSTO-211H cells) demonstrated significant growth Disclosure: No significant relationships. inhibition at doses less than 1.6 μM.

Conclusion: Our study suggests that inhibition of MTOR pathway may be an important therapeutic strategy in patients with MPM. Resistance POSTER SESSION 5 September 14, 2012 11:30-12:30 to Cetuximab may be overcome by combining Cetuximab with Celecoxib or MTOR kinase inhibitors (if PTEN is lost). Lack of cytotoxic effect with P5.04: A DUAL NFKB/STAT3 INHIBITOR AFFECTS Gefitinib may be due to the absence of EGFR mutations. CHEMORESISTANCE OF MALIGNANT PLEURAL MESOTHELIOMA CELLS. Disclosure: No significant relationships. Claudia Canino, Chandra Goparaju, J S. Donington, Harvey Pass Cardiothoracic Surgery, Nyu Langone Medical Center, New York/NY/ UNITED STATES OF AMERICA

Background: Chronic inflammation is an active mechanism of neoplastic progression. NFκB and STAT3 translate microenvironmental stimuli into inflammatory signaling. NFκB and STAT3 are costitutively activated in many solid tumors and their activated status correlates with the acquisition of mesenchymal features and with radio- and chemoresistance, hallmarks of aggressive tumors. The Epithelial-to- Mesenchymal transition and extreme chemoresistance are major features of the Malignant Pleural

iMig2012.org • Abstract Book 146 Mesothelioma in vivo and in vitro, and constitutive activation of NFκB has Results: In co-cultures, primary fibroblasts or fibroblast-conditioned been shown in asbestos treated mesothelial cells. Moreover, high levels medium induced activation of the pathways known to contribute to of nuclear, phosphorylated STAT3 have been observed in a conspicuous malignant transformation. Although subcellular location of damage- fraction of MPM specimens. We have shown that STAT3 activation is associated molecular pattern (DAMP) protein HMGB1 remained nuclear in required for the maintenance of a pool of chemoresistant, Mesothelioma- normal mesothelial cells co-cultured or treated with fibroblast-conditioned Tumor-Initiating Cells endowed with high levels of aldehyde deidrogenase medium, the levels of growth modulators were altered in these cells. activity (ALDHbright). We have therefore tested whether double inhibition of both NFkB and STAT3 activation can affect the tumorigenic properties of Conclusion: Non-mesothelial cells instigate alteration of cellular signalling Mesothelioma cell lines and the ALDHbright cell number. in mesothelial cells. Further integral examination of the aberrant signalling pathways, especially at early stages of neoplasia, will contribute to the Methods: We have used indirect immunofluorescence, WB, and protein development of more effective therapeutic strategies and in the longer immunoprecipitation to unravel a physical interaction of the NfKB and term may lead to personalised therapeutic approaches. STAT3 in MPM cell lines. We have then tested the effect of butein on the levels of EMT transcripts by Q-PCR and on the viability, clonogenicity, and Disclosure: No significant relationships. invasive properties of MPM cell lines. Finally, we have tested the ability of the drug to affect the number of ALDHbright cells and to overcome resistance to Pemetrexed in vitro and in vivo, by mean of FACS analysis and xenograft transplantation assays. POSTER SESSION 5 September 14, 2012 11:30-12:30 poster sessions | september 14

Results: We show here that multiple MPM cell lines contain high levels P5.06: ASBESTOS FIBERS INDUCE AN INFLAMMATORY of nuclear NfKB and pSTAT3(Y705). We also show that the two proteins RESPONSE THROUGH THE ADIPOCYTES BY DYSREGULATED interact in nuclear lysates. Treatment of MPM cells with naturally occurring ADIPOCYTOKINE PRODUCTION NFkB/STAT3 inhibitor (butein, 3,4,2’,4’-tetrahydroxychalcone) affects the STAT3 tyrosine phosphorylation and disrupts the NFkB-pSTAT3 interaction. Shan Hwu Chew1, Yasumasa Okazaki2, Shinya Toyokuni2 This correlates with a downregulation of several genes involved in cancer 1Pathology And Biological Responses, Nagoya University, Nagoya/ progression (such as ICAM1, Vimentin, MMP9, Twist), of proangiogenic JAPAN, 2Nagoya University/JAPAN cytokines (VEGF) and of IL-6 and IL-8, key growth factors for MPM. Additionally, butein affects the clonogenicity, invasion and the resistance Background: Asbestos-induced carcinogenesis is closely associated to pemetrexed of treated MPM cells. We show that these effects may be with a state of chronic inflammation. Asbestos exposure always results mediated by a significant reduction of the ALDHbright chemoresistant in an inflammatory response and previous evidence has indicated the cell subpopulations in butein-treated MPM cells. Interestingly, Butein is involvement of macrophages and mesothelial cells in such inflammatory effective on unsorted cell cultures rather than on FACS sorted ALDHbright response through production of pro-inflammatory molecules when they cells, suggesting the interference with paracrine signaling between encounter asbestos fibers. Those studies have highlighted the crucial the ALDHbright cells and the other cell subpopulations. In vivo, butein role of pro-inflammatory molecules as potential tumor promoter in the treatment severely affects tumor engraftment and potentiates the process of asbestos-induced carcinogenesis. Previously, several groups anticancer effects of Pemetrexed in mouse xenograft models. Interestingly, have argued that intraperitoneal injection is a better option to assess Butein does not significantly affect the viability of human, untransformed carcinogenic effect of fibers compared to intrapleural route as peritoneal mesothelial cells in vitro, nor does it affect survival of tumor-free mice in cavity was more sensitive to the effect of fibers. We hypothesized that vivo. the presence of abundant adipose tissue in the peritoneal cavity might underlie such higher sensitivity as there is a higher chance for direct Conclusion: We suggest that Butein may represent the prototype of a contact between adipocytes and asbestos fibers. Moreover, there has novel dual NFkB-STAT3 inhibitor of potential therapeutic interest for MPM. been tremendous evidence on the involvement of adipose tissue in inflammation through its ability to secrete various pro-inflammatory/ Disclosure: No significant relationships. anti-inflammatory molecules collectively known as the adipocytokines. Our international mesothelioma interest group study was aimed to investigate the ability of asbestos fibers to induce an inflammatory response from adipocytes, particularly through dysregulation of adipocytokine production. POSTER SESSION 5 September 14, 2012 11:30-12:30 Methods: We exposed differentiated mouse 3T3-L1 adipocytes to different P5.05: PRO-TUMORIGENIC ALTERATION OF SIGNALLING types of asbestos fibers followed by microarray analysis to identify genes PATHWAYS IN NORMAL MESOTHELIUM: CONTRIBUTION OF which are significantly upregulated or downregulated. qPCR and ELISA NON-MESOTHELIAL CELLS were performed to confirm the upregulation of a gene at mRNA and protein level. Several in vitro assays such as transwell migration assay, cell viability Tatyana Chernova1, Stefano Grosso1, Xiao Ming Sun1, Pavithra Kumar1, assay and cell proliferation assay were also performed. We also injected Fiona Murphy1, Jonathan Bennett2, Apostolos Nakas2, Martin Bushell1, asbestos fibers into the peritoneal cavity of animals to assess whether Marion Macfarlane1, Anne E. Willis1 similar changes could be observed in the adipose tissue in vivo. 1Toxicology Unit, Mrc, Leicester/UNITED KINGDOM, 2Glenfield Hospital, University Hospitals Of Leicester Nhs Trust, Leicester/UNITED KINGDOM Results: An adipocytokine, MCP-1 was significantly upregulated in 3T3-L1 adipocytes following asbestos exposure. qPCR and ELISA confirmed the Background: Malignant mesothelioma (MM) is an aggressive, fatal tumour increased mRNA level and secretion of MCP-1 protein into culture medium of the pleura or peritoneum and strongly related to asbestos exposure. respectively. We hypothesized that the increased MCP-1 production by the Malignant pleural mesothelioma (MPM) is the most common and occurs adipocytes in response to asbestos exposure can potentially contribute with a latency of up to 40 years. The mechanism of MM carcinogenesis is to asbestos-induced carcinogenesis via two mechanisms: enhanced not well understood and the heterogeneity of the tumour is considered to macrophage recruitment to sites of asbestos deposition as well as direct be a major barrier to successful therapies. Several studies have identified stimulation of mesothelial cell proliferation. Conditioned medium from changes in the expression and activities of defined cell signalling pathways adipocytes treated with asbestos fibers has a higher ability to induce in mesothelial and stromal cells, but the relationship between different cell macrophage chemotaxis, which is in agreement with increased secretion of types in the process of tumorigenesis has not been studied. MCP-1 protein into the culture medium. Treatment of mesothelial cells with recombinant MCP-1 resulted in enhanced cell proliferation. Examination Methods: To examine the pro-oncogenic role(s) of different cell of MCP-1 expression in the adipose tissue of mice injected with asbestos populations, the effect of activated fibroblasts and macrophages on cellular fibers further supports the in vitro findings. Interestingly, we also noted signalling in normal untransformed mesothelial cells was monitored using suppressed level of an anti-inflammatory adipocytokine, adiponectin, in imaging and immunoblotting techniques. the adipose tissue of mice following asbestos exposure.

iMig2012.org • Abstract Book 147 Conclusion: We have shown that asbestos fibers were able to directly POSTER SESSION 5 September 14, 2012 11:30-12:30 stimulate an inflammatory response from the adipoctyes, which occurs through dysregulated adipocytokine production. Increased MCP-1 P5.08: CURCUMIN KILLS MALIGNANT MESOTHELIOMA CELLS BY production might have an important implication as it might indirectly PYROPTOSIS aggravate the inflammation associated with asbestos fibers through recruitment of macrophages or directly stimulate the proliferation of Jill M. Miller, Maximilian Macpherson, Stacie Beuschel, Mutlay Sayan, mesothelial cells. Either way, our results suggest that the adipocytes can Brooke T. Mossman, Arti Shukla potentially act as a tumor promoter in asbestos-induced carcinogenesis Pathology, University Of Vermont, Burlington/UNITED STATES OF AMERICA through dysregulated adipocytokine production. Background: Malignant mesothelioma (MM) is an asbestos assoicated Disclosure: No significant relationships. maligancy with a dismal progrosis and poor therapeutic strategies. Curcumin, a natural occuring polyphenol in turmic, has been shown to have anticarcinogenic properties in multiple cancers, however little research has explored the therapeutic role of curcumin in MM. We POSTER SESSION 5 September 14, 2012 11:30-12:30 hypothesized that curcumin would reduce proliferation and growth of MM via pyroptosis, an inflammation-mediated cell death process dependent on P5.07: THE MESOTHELIOMA TUMOR SUPPRESSOR BAP1 inflammsome activation of caspase-1. INTERACTS WITH PRKDC, PART OF THE DNA-PK DNA DAMAGE poster sessions | september 14 REPAIR PROTEIN COMPLEX. Methods: For our experiments, we used telomerase immortalized human peritoneal mesothelial cells (LP9) and mouse MM cells (#40). Cell growth Tatsuo Ito1, Hediye E. Bromage2, Marc Ladanyi3 studies were perfromed by MTT assays. Steady-state mRNA levels of 1Department Of Pathology, Memorial Sloan Kettering Cancer Center, New NOD-like receptor protein 3 (NLRP3), High Mobility Group Box 1 (HMGB-1), York/NY/UNITED STATES OF AMERICA, 2Microchemistry And Proteomics and pro-IL-1β in LP9 and mouse MM cells was assessed by qRT-PCR using Core, Memorial Sloan Kettering Cancer Center, New York/NY/UNITED specific primers and probes. Caspase-1 activity, after various treatments STATES OF AMERICA, 3Pathology And Human Oncology & Pathogenesis with curcumin and/or asbestos, was evaluated by the Caspase-1 Program, Memorial Sloan-Kettering Cancer Center, New York/NY/UNITED Colorimetric Assay. HMGB-1 in supernatants was measured by Western blot STATES OF AMERICA analysis. IL-1β in supernatants was measured by ELISA.

Background: Recent studies (Bott et al., Nat Genet 43:668-72, 2011) have Results: LP9 cells treated with curcumin 10 µM for 24-72 hours and identified frequent inactivation of the tumor suppressor gene BAP1 (BRCA exposed to asbestoes for 24-48 hours had increased NLRP3 mRNA levels associated protein 1) in malignant pleural mesothelioma (MPM). and showed a trend in increased caspase-1 and IL-1β activity compared to Additionally, germline mutations in BAP1 have been identified and an control and asbestos exposed LP9 cells. MTT assay results revealed that associated cancer syndrome, which includes MPM, ocular melanoma and curcumin inhibited mouse MM cell growth in a dose and time dependent other cancers, has been described (Testa et al., Nat Genet 43:1022-5). fashion. Mouse MM cells treated with curcumin 40 µM for 48 hours had BAP1 includes an N-terminal ubiquitin C-terminal hydrolase (UCH) domain, largely significant increases in steady-state mRNA levels of NLRP3, pro-IL- and a nuclear localization signal (NLS) at the C-terminal end. The majority 1beta, and HMGB-1. Additionally, these cells had significantly increased of BAP1 truncating mutations are predicted to result in loss of the nuclear caspase-1 activity and HMGB-1 present in supernatants compared to localization signal and/or the C-terminal UCH domain. How BAP1 functions control mouse MM cells. as a tumor suppressor is still unclear. Here, we tried to identify interaction partners of BAP1 in MPM cells and to begin to define functional aspects of Conclusion: Our in vitro data indicates that curcumin is able to suppress these interactions. MM cell growth through pyroptosis as demonstrated by increased steady- state levels of NLRP3 inflammasome, caspase-1 activation and release of Methods: We used protein co-immunoprecipitation, mass-spectroscopy HMGB-1, a damage associated molecular protein released in response to analysis, ubiquitin AMC assays, and immunostaining to identify and analyze activation of caspase-1. A similar trend in inflammasome transcription and international mesothelioma interest group BAP1 interaction partners. Single cell gel electrophoresis (“Comet”) assays activation was observed in LP9 cells pretreated with curcumin and exposed were performed to assess DNA damage in MPM cells with and without BAP1 to asbestos. These results provide evidence that curcumin warrants further protein expression. investigation as a potential therapeutic agent in MM. We plan to validate our findings using an in vivo mouse MM model in which we will evaluate the Results: Mass-spectroscopy analysis of BAP1 protein complexes in H-Meso ability of curcumin alone and in combination with other drugs to suppress MPM cells and 293T cells (human embryonic kidney) identified known intraperitoneal mouse MM tumor development. This work is supported binding partners [HCF1, ASXL2, histone 2A (H2A)] as well as possible novel by Mesothelioma Applied Research Foundation (MARF) grant (AS) and by interaction partners, notably PRKDC which encodes the catalytic subunit NIEHS grants 1RO1ES021110 (AS), T32 ES07122(BM). of the DNA-dependent protein kinase (DNA-PK) and functions with the Ku70/Ku80 heterodimer in DNA double strand break repair. Binding of Disclosure: No significant relationships. BAP1 to PRKDC was mediated by the C-terminal portion of BAP1. DNA-PK inhibitor II (Calbiochem) reduced the activity of BAP1 as a deubiquitinase in vitro, based on the ubiquitin AMC assay. In Comet assays, BAP1 knockdown was associated with increased DNA damage (increased tail length) and this POSTER SESSION 5 September 14, 2012 11:30-12:30 effect was further enhanced by concomitant DNA-PK inhibitor II treatment. P5.09: COMBINATORIAL APPROACHES FOR TARGETING Conclusion: BAP1 interacts with DNA-PK protein complexes involved in MITOCHONDRIAL REDOX SIGNALING AND FOXM1 EXPRESSION IN DNA damage repair. We hypothesize that phosphorylation of BAP1 by MALIGNANT MESOTHELIOMA PRKDC may be important to activate the de-ubiquitination activity of BAP1. These data may point to new targetable key pathways in MPM. Kheng Newick1, Brian Cunniff1, Balaraman Kalyanaraman2, Paul Held3, Jack Arbiser4, Harvey Pass5, Brooke T. Mossman1, Arti Shukla1, Nicholas Disclosure: No significant relationships. Heintz1 1Pathology, University Of Vermont, Burlington/UNITED STATES OF AMERICA, 2Department Of Biophysics And Free Radical Research Center, Medical College Of Wisconsin, Milwaukee/UNITED STATES OF AMERICA, 3Biotek Instruments, Winooski/UNITED STATES OF AMERICA, 4Dermatology, Emory University, Atlanta/UNITED STATES OF AMERICA, 5Cardiothoracic Surgery, New York University Medical Center, New York/NY/UNITED STATES OF AMERICA

iMig2012.org • Abstract Book 148 Background: Redox-dependent signaling by reactive oxygen species (ROS) plays an important role in cancer pathogenesis, and may represent a therapeutic target in malignant mesothelioma (MM). We have explored the role of FOXM1, a redox-responsive forkhead transcription factor that regulates cell cycle progression and resistance to oxidative stress, in MM cell proliferation and viability.

Methods: The detection of mRNA and protein were performed using qPCR arrays and western blotting, along with immunohistochemistry and microscopy techniques. The detection of reactive oxygen species (ROS) was performed using ROS probes, and the effects of our redox-sensitive drugs were tested in an in vivo model where a xenograft model of human mesothelioma cells were inoculated into severe combined immunodeficient (SCID) mice. Interestingly, TP53 polymorphisms in intron 7 (rs12947788 and rs12951053) Results: Human MM tumors express more FOXM1 transcript than were identified in both asbestos-related thoracic cancers. normal mesothelial tissue, and immunostaining of human MM tissue arrays confirms that FOXM1 is broadly expressed in all major subtypes of poster sessions | september 14 human MM. Studies show that MM cells in vitro constitutively generate approximately 2-fold more mitochondrial superoxide than control immortalized LP9 mesothelial cells. The triphenylmethane gentian violet (GV), the thiazole antibiotic thiostrepton (TS), and selected triphenylphosphonium (TPP) compounds inhibit FOXM1 expression and MM tumor cell viability in a dose–dependent manner. We have shown each of these agents targets different facets of the thioredoxin reductase 2 (TR2) - thioredoxin 2 (TRX2) - peroxiredoxin 3 (PRX3) antioxidant network, the predominate pathway for metabolizing hydrogen peroxide in mitochondria. GV targets expression of TRX2 protein, TS covalently adducts and inactivates PRX3, and TPP compounds inhibit expression of PRX3 and induce extensive mitochondrial fragmentation. In contrast, the general NADPH oxidase inhibitor DPI had no effect on FOXM1 expression. Studies Disclosure: No significant relationships. in a xenoplant model of human MM in Fox Chase SCID mice show that TS and GV alone are marginally effective in inhibiting tumor growth, while a combination of the two compounds significantly impairs tumor progression without overt toxicity (p = 0.006). Our in vitro studies indicate that FOXM1 POSTER SESSION 5 September 14, 2012 11:30-12:30 expression is tuned to an optimal flux of mitochondrial oxidant production, and agents that either markedly diminish or accentuate the production of P5.11: EPHRINA1 MEDIATED OVER EXPRESSION OF MICRORNA- mitochondrial oxidants inhibit FOXM1 expression. 302B INDUCES APOPTOSIS IN MALIGNANT MESOTHELIOMA

Conclusion: Since over-expression of the TR2-TRX2-PRX3 pathway is Nazli Khodayari1, Kamal A. Mohammed1, Eugene P. Goldberg2, Michael linked to resistance to apoptosis, and components of the pathways are A. Jantz1, Najmunnisa Nasreen1 known to be up-regulated in the majority of MMs, these studies offer 1Medicine, University Of Florida, Gainesville/FL/UNITED STATES OF a new therapeutic strategy for treating MM. Moreover, inhibition of AMERICA, 2Biomaterial Science And Engineering, University Of Florida, PRX3 expression has been shown to sensitize cancer cells to common Gainesville/FL/UNITED STATES OF AMERICA international mesothelioma interest group chemotherapeutic drugs, suggesting that alone or in selected combinations these agents may be useful in disabling a major adaptive response that Background: EphrinA1-EphA2 signaling control variety of cellular functions contributes to MM drug resistance. Generously supported by the John including proliferation, migration, invasion and apoptosis. EphrinA1 Sterling Family Memorial Grant from the Mesothelioma Applied Research activation of receptor EphA2 induces anti-tumorogenic effects in malignant Foundation, the Lake Champlain Cancer Research Organization, the Ladies mesothelioma (MM). In addition there is growing evidence that ephrinA1 Auxiliary of the Vermont VFW, and the Vermont Cancer Center. induces microRNAs expression in malignant cells. MicroRNAs are key players in the control of cell proliferation and cell fate determination. However, Disclosure: No significant relationships. the molecular mechanisms associated with ephrinA1 and receptor EphA2 mediated attenuation of MM tumor growth remains unclear. Our study identified a novel mechanism of ephrin-A1 mediated tumor suppressor signaling in MM. Ephrin-A1 activation up regulates miR-302b expression in POSTER SESSION 5 September 14, 2012 11:30-12:30 MM and attenuates tumor growth via repression of myeloid cell leukemia (Mcl-1), an anti-apoptotic protein of the Bcl-2 family. P5.10: DIFFERENTIAL TP53 MUTATIONS AND INTRONIC POLYMORPHISMS IN ASBESTOS-RELATED LUNG CANCER AND Methods: MM cell lines (CRL-2081=MMC1; and CRL-5830=MMC2) were MALIGNANT PLEURAL MESOTHELIOMA. grown to near confluence, and activated with recombinant-Ephrin-A1- Fc for the indicated period of time. The expression of miR-302b was Pascal Andujar1, Jean-claude Pairon1, Annie Renier2, Alexis Descatha3, analyzed by Microarray, quantitative real time RT-PCR analysis. Mcl-1 and Françoise Galateau-Sallé4, Françoise Le Pimpec-Barthes2, Jessica cell cycle gene expression were analyzed by RT-PCR, immuno-blotting Zucman-Rossi2, Marie-Claude Jaurand2, Didier Jean2 and Immunofluorescence analysis. To confirm that ephrin-A1 regulates 1Service De Pneumologie Et Pathologie Professionnelle, Centre Hospitalier the expression of Mcl-1 mRNA through miR302-b up regulation, cultured Intercommunal De Créteil, Créteil/FRANCE, 2Umr-S 674, Inserm - Université cells were transfected with and without miR302-b precursors or miR- Paris Descartes, Paris/FRANCE, 3U1018, Inserm, Villejuif/FRANCE, 4Service 302b inhibitor prior to activation. The tumor growth was measured by 3D D’Anatomie Pathologique, Chu De Caen, Caen/FRANCE matrigel assay. MM apoptosis was determined by TUNEL assay and Flow cytometry. MiR-302b binding to the 3’UTR of human Mcl-1 was confirmed Results: TP53 mutations were found in NSCLC with no link with asbestos by Luciferase assay. exposure and the frequency was higher in NSCLC than in MPM. However, significant enhancement of TP53 G:C to T:A transversions was found in Results: Ephrin-A1 activation induced several fold increases of miR-302b NSCLC from asbestos-exposed patients. expression in MM cells when compared to respective untreated MM cells.

iMig2012.org • Abstract Book 149 Ephrin-A1 activation significantly down regulated Mcl-1 expression in MM Conclusion: Glibenclamide sensitizes MPM cell lines and primary cells compared to resting cells. Moreover, ephrin-A1 activation significantly cultures to TRAIL-mediated apoptosis, probably through different inhibited MM tumor growth. EphrinA1 activation induced 28.98% ± mechanisms of action in the epithelioid and sarcomatoid histotypes; in 2.38 ( p <0.05), and 22.26% ± 2.74; (p <0.05) cell death in MMC1 and fact a ROS-dependent p53 activation was observed in epithelioid but MMC2 respectively. The transfection of MMC with miR302b inhibitor and not in sarcomatoid cell lines. Further studies should test the efficacy of activation with ephrinA1 significantly restored the expression of Mcl-1. In Glibenclamide in the treatment of MPM patients. addition transfection of MM cells with miR302-b vector induced apoptosis. Furthermore, the cell cycle genes cyclin D1, and D2 of G1-S phase were Disclosure: No significant relationships. affected indicating that ephrinA1 activation of receptor EphA2 controls the fate of MMC through miR302b expression and induces apoptosis.

Conclusion: We have identified a novel mechanism of ephrinA1 mediated POSTER SESSION 5 September 14, 2012 11:30-12:30 signaling in MM. Our results suggest that in MM ephrinA1 by inducing miR302b expression attenuates tumor growth. MiR-302b targets Mcl- P5.13: TUMOR-ASSOCIATED MACROPHAGES MEDIATE 1 gene and also affects the expression of cell cycle genes and thereby RESISTANCE TO PEGYLATED ARGININE DEIMINASE IN induces apoptosis. These findings suggest that ephrinA1 could be a MALIGNANT PLEURAL MESOTHELIOMA promising therapeutic agent for the treatment of MM. Melissa Phillips1, Kevin Sharpe2, Jeremy P. Steele3, Fiona Mccarthy4, poster sessions | september 14 Disclosure: No significant relationships. Laura Tookman1, Fiona Luong2, Thorsten Hagemann4, Peter Szlosarek2 1Molecular Oncology And Imaging, Barts Cancer Institute, Queen Mary University Of London, London/UNITED KINGDOM, 2Molecular Oncology, Barts Cancer Institute, Queen Mary University Of London, London/UNITED POSTER SESSION 5 September 14, 2012 11:30-12:30 KINGDOM, 3Medical Oncology, St Bartholomew’s Hospital, London/UNITED KINGDOM, 4Cancer And Inflammation, Barts Cancer Institute, Queen Mary P5.12: GLIBENCLAMIDE SENSITIZES MALIGNANT PLEURAL University Of London, London/UNITED KINGDOM MESOTHELIOMA CELLS AND PRIMARY CULTURES TO TRAIL- MEDIATED APOPTOSIS Background: Malignant pleural mesothelioma (MPM) deficient in argininosuccinate synthetase (ASS1), a rate-limiting enzyme in the Giulia Pasello1, Loredana Urso1, Micol Silic-Benussi2, Marco Schiavon3, biosynthesis of arginine, is sensitive to arginine deprivation. The discovery Vincenzo Ciminale2, Federico Rea3, Adolfo Favaretto1 of this ‘Achilles’ heel’ in approximately 50% of MPM has led to the initiation 1Medical Oncology, Istituto Oncologico Veneto, Padua/ITALY, 2Surgical of a UK wide randomised trial of the arginine depleting agent pegylated Oncologic And Gastroenterological Sciences Dept, University Of Padua, arginine deiminase (ADI-PEG20) in patients with MPM, following on from Padua/ITALY, 3Thoracic Surgery Department, University Of Padua, Padua/ promising earlier trials in patients with other arginine dependent tumors. ITALY However, tumoral resistance to arginine depletion has been observed: although several mechanisms have been shown to mediate this, including Background: Malignant pleural mesothelioma (MPM) is an aggressive up-regulation of ASS1, autophagy and drug-neutralising antibodies, the tumour with poor prognosis and increasing incidence in industrialized supply of arginine specifically by ASS1-rich stromal cells remains to be countries. MPM is highly refractory to current chemotherapy, and complete defined. Here, we hypothesised that tumor-associated macrophages responses are rarely observed. To improve MPM therapy we tested the (TAMs), especially abundant in MPM, may act as ‘feeder cells’ providing synergic effect of engaging the Tumor necrosis factor-Related Apoptosis- MPM with arginine (or its intermediates), thereby bypassing the effect of Inducing Ligand (TRAIL) apoptotic pathway and Glibenclamide, an arginine depletion. antidiabetic drug which showed a Reactive Oxygen Species (ROS) inducing effect. Methods: Three ASS1 negative MPM cell lines treated with and without ADI-PEG20 were analysed using the Affymetrix U133 plus 2.0 microarray international mesothelioma interest group Methods: Apoptosis assays were performed using Fluorometric platform and validated using qPCR and ELISA. Monocytes were isolated Homogenous Caspase Assay (Roche). MPM cells were previously treated from human buffy cones using CD14 positive selection and matured in with or without TRAIL (50 ng/ml) and/or Glibenclamide (50 uM) for 24 2% human AB serum for seven days. Macrophage influence on tumoral hours and then incubated with DEVD-Rhodamine 110. Upon cleavage of the arginine supply was assessed in vitro by culturing macrophages both substrate by activated caspases, fluorescence of the released Rhodamine with and without direct MPM cell contact in the presence and absence 110 was measured by a microplate reader. ROS assays were performed of ADI-PEG20. Tumor cell viability was determined by day four using flow using MitoTracker Red CM-H2XRos, which accumulates in mitochondria cytometry. Macrophage ASS1 expression from co-culture was assessed

and produces a red fluorescent signal upon oxidation by H2O2. MPM cells by qPCR and western blot. Levels of argininosuccinate in the supernatant were pre-incubated with 25nM H2-MTR, then treated with Glibenclamide were measured using HPLC. ADI-PEG20 in combination with macrophage and fluorescence was analyzed every 5 minutes for at least 45 minutes depletion (using liposomal clodronate (CLIP)) was assessed in vivo using an using a Zeiss LSM510 confocal laser microscope. p53 protein expression MPM xenograft model. level was detected by western blot analysis using a specific antibody. Results: Distinct immune and inflammatory genes were modulated in all Results: Caspases activity in 8 cell lines (4 epithelioid, 1sarcomatoid, three ASS1 negative cell lines using Affymetrix gene expression analyses. In 3 biphasic) and one primary culture was analyzed after treatment with particular, the mRNA and protein of several cytokines involved in myeloid Glibenclamide, TRAIL, Glibenclamide + TRAIL. We observed a statistically cell recruitment/activation, including VEGFA, IL8, CXCL2, CXCL3, FGF2 significant increase of caspases activity in all the cell lines treated with and IL1alpha, were significantly up-regulated by 24 hours in the ADI- the combination of the two agents compared to untreated controls and PEG20 treated ASS1 negative cell lines, compared with control. Co-culture to TRAIL and Glibenclamide used as single agents. We analyzed ROS levels results revealed a 10% increase in MPM cell viability in cells cultured with in two cell lines (epithelioid ZL55 and sarcomatoid ZL34); we observed macrophages without direct cell contact, increasing up to 30% when co- ROS induction in ZL55 treated with Glibenclamide + TRAIL compared to cultured with macrophages in direct cell contact. There was a significant no treatment, while no higher ROS levels were assessed in ZL34 treated increase in ASS1 expression in co-cultured macrophages 48 hours after with the two agents compared to untreated. Since it was demonstrated treatment with ADI-PEG20, compared with macrophages cultured that ROS can lead to p53 activation, we examined whether Glibenclamide alone. Initial HPLC results have shown that levels of argininosuccinate in treatment could activate p53 protein in ZL55 and ZL34 cell lines. As supernatant from co-cultured macrophages are significantly higher than expected, treatment with Glibenclamide increased p53 activation only in from control macrophages. Preliminary results from ongoing xenograft ZL55 cells and this activation was reverted by pre-treatment with the ROS studies have demonstrated a significant reduction in tumor volume in mice scavenger N-acetyl-cysteine (NAC). treated with ADI-PEG20 in combination with CLIP, compared with single agent ADI-PEG20.

iMig2012.org • Abstract Book 150 Conclusion: Collectively, these results indicate that TAMs may modulate can be more reliably modeled. These findings highlight the need for the sensitivity to ADI-PEG20 in MPM by protecting the tumor cells from development of more appropriate and well-characterized models of MPM. arginine deprivation via the provision of arginine or its substrates. Future work aims to interrogate specific pro-inflammatory pathways to assess Disclosure: No significant relationships. whether this TAM-mediated drug resistance is reversible.

Disclosure: No significant relationships. POSTER SESSION 5 September 14, 2012 11:30-12:30

P5.15: M14K AND M38K MALIGNANT PLEURAL MESOTHELIOMA POSTER SESSION 5 September 14, 2012 11:30-12:30 CELL LINES CONSERVE BOTH IN VIVO AND IN VITRO THE SAME CLAUDIN- BASED PHENOTYPE P5.14: COMMONLY USED MESOTHELIOMA CELL LINES EXHIBIT LOSS OF MESOTHELIAL MORPHOLOGY AND MARKERS, Siham Chaouche-Mazouni1, Marie-Christine Copin2, Nemcha BUT RETAIN MICRORNA EXPRESSION PATTERNS OF NATIVE Lebaili1, Philippe Hauw3, Corine Glineur3, Anne Tsicopoulos3, Philippe TUMOURS Lassalle3, Arnaud Scherpereel4 1Ecole Normale Supérieure (Ens) Of Kouba, Algiers/ALGERIA, 2Pôle Glen Reid1, Kim Griggs2, Michaela B. Kirschner3, Yuen Y. Cheng1, Biologie Pathologie, Chru De Lille, Lille/FRANCE, 3Center For Infection poster sessions | september 14 J.J.B. Edelman4, Michael P. Vallely4, Brian C. Mccaughan4, Douglas And Immunity Of Lille, Inserm U1019-Cnrs 8204, Institut Pasteur De Lille/ Henderson5, Nico Van Zandwijk1, Sonja Klebe2 FRANCE, 4Pulmonary And Thoracic Oncology, Lille University Hospital, 1Asbestos Diseases Research Institute, Concord/NSW/ Lille/FRANCE AUSTRALIA, 2Department Of Anatomical Pathology, Filnders Univerity, Adelaide/AUSTRALIA, 3Asbestos Diseases Research Institute, University Of Background: Malignant Pleural Mesothelioma (MPM) is a rare and Sydney, Concord/AUSTRALIA, 4Cardiothoracic Surgical Unit, Royal Prince aggressive neoplasm with difficult diagnosis, and poor prognosis. Cell lines Alfred Hospital; The Baird Institute And Faculty Of Medicine, University Of available for mesothelioma research are rare and often heterogeneous. Sydney, Newtown/AUSTRALIA, 5Department Of Anatomical Pathology, Sa Claudins, a major component of tight junctions have been shown to be Pathology And Flinders University, Adelaide/AUSTRALIA novel, highly attractive targets for diagnosis and therapy of epithelial tumors. The aim of this study was to establish a claudin-based MPM Background: Malignant pleural mesothelioma (MPM) often responds poorly phenotype and to verify whether it is conserved in vitro and in vivo, to to treatment. The identification of new therapeutic approaches requires select the most representative MPM cell lines. the use of in vitro models, and many studies use established cell lines from the ATCC. In the literature, these cells lines are variously described Methods: The expression of claudin-3 and claudin-4 as well as calretinin as epithelioid, biphasic or sarcomatoid mesothelioma and occasionally was examined by immunohistochemistry and immuno-blot in fourteen non small cell lung cancer. Although these cell lines were isolated from human specimens of MPM (n=5), lung adenocarcinoma (ADC) (n=5) and mesothelioma patients, they have not been extensively characterized with normal lung tissues (n=4). The expression of these proteins in pathological respect to their growth characteristics, their electron microscopic features tissues was compared, in addition to claudin-15 protein with that in human and expression of surface markers characteristic to mesothelial cells -- MPM-derived cell lines (i.e. MSTO-211, NCI-H28, M38K and M14K), in features used in the clinical diagnosis of MPM. human ADC-derived cell lines (i.e. A549, HT29, MCF7 and MDA-MB-231) and finally in human MPM or ADK-derived subcutaneous tumor xenografts Methods: The immortalized human mesothelial cell line MeT-5A and obtained in SCID mice. human mesothelioma cell lines H28 (mesothelioma NOS), NCI-H226 (epithelial) and MSTO-H211 (biphasic) were obtained from the ATCC and Results: A well defined MPM phenotype based on the expression of cultured as recommended. In addition, spheroid (3D) cultures were also claudin-15 and calretinin and the absence of expression of claudin-3 grown. Subcutaneous xenografts were generated as described previously. and claudin-4 was established. We found claudin-15 as well as calretinin international mesothelioma interest group Mesothelial marker and markers expected to be negative in MPM were highly expressed by M14K and M38K whereas claudins-3 and -4 were not assessed by immunohistochemistry (IHC) of cultured cells and xenograft expressed in these two MPM cell lines. Identical results were obtained in tumours. Structural morphology of cultured cells was assessed by SCID mice cell line-derived tumor xenografts. transmission electron microscopy (TEM). RNA was isolated from cells in 2D and 3D culture and xenografts, and the expression of microRNAs was Conclusion: Our results suggest that M14K and M38K cell lines in vitro assessed using TaqMan low-density arrays. or in tumor xenografts in SCID mice are representative models that could be used to improve diagnosis and prognosis tools and to explore new Results: IHC revealed that the characteristic mesothelial markers therapeutic strategies in MPM targeting claudins. calretinin and D240 were expressed by few cells in cultures of MeT-5A, H28, H226 and MSTO-H211. These expression profiles altered slightly in Disclosure: No significant relationships. the in vivo tumours. In contrast, CD15 and BG8 are carcinoma-markers characteristically negative in normal and neoplastic mesothelium and used as negative markers for mesothelioma clinically, but were expressed in some of the cell lines: H28 showed marked expression of CD16, and MeT- POSTER SESSION 5 September 14, 2012 11:30-12:30 5A showed weaker focal expression of both CD15 and BG8. TEM showed that mesothelial cell characteristics were frequently absent, including from P5.16: IDENTIFICATION OF CELL DEATH PATHWAYS THAT ARE the ‘normal’ mesothelial cell line MeT5A, with few microvilli, intermediate DIFFERENTIALLY DEREGULATED IN MALIGNANT PLEURAL filaments or glycogen observed, and no evidence of intercellular junctions. MESOTHELIOMA Of the mesothelioma cell lines assessed, H226 most closely resembled mesothelial-like cell morphology, whereas H28 had the least in common Xiao Ming Sun1, Tatyana Chernova1, Stefano Grosso1, Fiona Murphy1, with native tumours. In contrast to the loss of characteristic mesothelial Jonathan Bennett2, Apostolos Nakas2, Martin Bushell1, Kelvin Cain3, cell marker expression and morphology, changes in microRNA expression Anne E. Willis1, Marion Macfarlane1 between MeT-5A and mesothelioma lines more closely paralleled changes 1Toxicology Unit, Mrc, Leicester/UNITED KINGDOM, 2Glenfield observed between normal mesothelium and tumour specimens. Hospital, University Hospitals Of Leicester Nhs Trust, Leicester/UNITED KINGDOM, 3Mrc Toxicology Unit, Leicester/UNITED KINGDOM Conclusion: The cell lines most commonly used to study MPM and test novel treatment regimens have retained few of the morphological Background: Malignant mesothelioma (MM) is an aggressive, fatal tumour attributes characteristic of mesothelial cells, limiting their suitability for of the pleura or peritoneum and is strongly related to asbestos exposure. some in vitro functional studies. MicroRNA expression, on the other hand, Malignant pleural mesothelioma (MPM) is the most common form of

iMig2012.org • Abstract Book 151 MM and occurs with a latency of up to 40 years. The mechanism of MM expression. We found by Ingenuity pathway analysis (IPA) that the most carcinogenesis is not well understood and to date, there is no curative affected functions were cellular movement, cell death, cellular growth therapy. Although the profound resistance of MM to cytotoxic agents is and proliferation, cellular signaling, development and cell cycle. Applying well documented, and is reportedly due to inhibition of cell death, the a network enrichment analysis approach, from more than 1,600 pathways identification of which cell death pathways are deregulated in this disease analyzed, 939 were significantly altered in syndecan-1 over-expressing has not been systematically examined. cells and 234 in syndecan-1 silenced cells. The results further expanded the findings from gene set enrichment analysis and IPA: many growth-factor, Methods: To obtain a more complete picture of which cell death cytokine and cell cycle -related pathways were altered: TGF-β, EGF, VEGF pathways are deregulated and to determine whether this is related to and ERK/MAPK pathways were enriched in both experimental settings. the aetiology of the disease we examined the expression of key cell death Nearly all analyzed pathways related to cell cycle were enriched after pathway regulators in different subtypes of human MPM cell lines, normal syndecan-1 silencing and depleted after syndecan-1 over expression. untransformed mesothelial cells and MPM tissue obtained from patients. In addition, we compared the sensitivity of normal mesothelial cells and Conclusion: Syndecan-1 seems to orchestrate different growth factors, MPM cell lines to a range of cytotoxic agents, including the DNA damaging converging to the downstream phosphokinase pathways. A better agents Cisplatin, Mitomycin C and Etoposide, as well as the highly selective understanding of the complex role of syndecan-1 and its molecular Bcl-2 antagonist, ABT 737. interactions may provide possibilities in the future to control growth of malignant mesothelioma. To address the individual contribution of the Results: Using this approach great heterogeneity was observed in altered pathways, functional studies are ongoing in our laboratory. poster sessions | september 14 the response of malignant cell lines to cytotoxic agents with profound resistance to cell death being evident in both epitheloid and sarcomatoid- Disclosure: No significant relationships. derived MPM cells; this in turn may account for the differing response of these tumours to treatment. Importantly, initial profiling of malignant mesothelioma tumour samples from patients has identified similar changes in several key cell death pathway regulators. POSTER SESSION 5 September 14, 2012 11:30-12:30

Conclusion: The role of these signalling pathways in disease development P5.18: STEROID RECEPTOR COACTIVATORS IN MALIGNANT and potential implications for future treatment of MPM will be discussed. PLEURAL MESOTHELIOMA: EXPRESSION AND PROGNOSTIC VALUE Disclosure: No significant relationships. Warren Thomas1, Cormac J. Jennings1, A O’Grady2, R Cummins2, B Murer3, E W Kay2, B J Harvey1 1Department Of Molecular Medicine, Royal College Of Surgeons In Ireland, POSTER SESSION 5 September 14, 2012 11:30-12:30 Dublin/IRELAND, 2Department Of Histology, Royal College Of Surgeons In Ireland/IRELAND, 3Department Of Pathology, Dell’Angello Hospital, P5.17: SIGNALING PATHWAYS INFLUENCED BY SYNDECAN-1 IN Mestre/ITALY MALIGNANT MESOTHELIOMA Background: Malignant pleural mesothelioma (MPM), which is associated Tünde Szatmári1, Filip Mundt1, Ghazal Heidari-Hamedani1, Fang Zong1, with asbestos exposure, is an aggressive disease arising from the pleural Elena Ferolla1, Andrey Alexeyenko2, Anders Hjerpe1, Katalin Dobra1 mesothelium. The incidence of MPM is higher in men than women which 1Department Of Laboratory Medicine, Karolinska Institute, Stockholm/ is probably linked to occupational exposure, however women experience SWEDEN, 2Science For Life Laboratory, Stockholm/SWEDEN longer post diagnosis survival. Recent studies indicate that this gender difference in survival may be due to hormonal status since expression of Background: The differentiation of malignant mesothelioma involves oestrogen receptor (ER)b is associated with better prognosis. Oestrogen syndecan-1, a cell surface heparan sulfate proteoglycan. Syndecan-1 signalling is amplified via the recruitment of steroid receptor coactivators international mesothelioma interest group modulates a number of interactions of different growth factors with their (SRCs) to the DNA bound steroid receptor. This study investigated the receptors, thus acting as a signaling co-receptor. We have previously association between clinical outcome and the expression of ERβ and the shown that syndecan-1 modulates malignant mesothelioma cell p160 group of SRCs. proliferation although the exact underlying molecular mechanisms are not completely elucidated. In this study we aim to characterize the molecular Methods: The expression of ERβ, SRC-1, SRC-2/TIF-2 and AIB-1/SRC-3 was events underlying the growth modulatory effect of syndecan-1 in malignant analysed by immunohistochemistry (IHC) in tumour biopsies from a cohort pleural mesothelioma and to identify critical factors and pathways of 89 confirmed MPM subjects and 3 control subjects. Allred scores were dependent on syndecan-1, focusing on cell-cycle regulation and features assigned based on expression of each of the proteins, and Kaplan-Meier related to proliferation. survival curves were generated. Western blot analysis was performed for each of the SRCs to compare receptor expression profiles between 8 MPM Methods: We modulated the expression of syndecan-1 in a human cell lines. mesothelioma cell line via both over expression and silencing and followed the transcriptomic responses with microarray analysis. To project Results: Normal mesothelium was highly positive for ERβ and each of the the transcriptome analysis on the full-dimensional picture of cellular SRCs, whereas variable expression levels were found in the MPM tumour regulation, we applied a novel method of network enrichment analysis samples. Kaplan-Meier survival analysis showed that ERβ expression or which elucidated signaling relations between differentially expressed genes high expression of TIF-2 (p = 0.036, log-rank test) conferred a survival and pathways acting via various molecular mechanisms. advantage across all mesothelioma cases. The normal mesothelium cell line Met-5A displayed high levels of expression for each of the SRCs while Results: Fourteen individual genes showed response to both up- and Y-Meso-8D and DM-3 sarcomatoid MPM cell lines displayed little or no down-regulation of syndecan-1. Syndecan-1 over expression had profound expression effects on genes involved in regulation of cell behavior whereas syndecan-1 silencing had less powerful effect, which can be explained by the already Conclusion: The improved survival for patients with ERβ expression and low initial syndecan-1 level of these cells. Syndecan-1 over expression high levels of TIF-2 expression may inform novel steroid-based approaches strongly affected cell proliferation: from 783 proliferation related genes on to therapeutic intervention for MPM. The use of specific ERβ agonists to the chip, 51 were downregulated and 74 were upregulated (p<0.001). The activate oestrogen signalling pathways in patients with MPM may offer an growth factor binding properties of syndecan-1 and its ability to regulate alternative therapeutic approach. cell growth depends on the fine structure of the heparan sulfate chains, particularly on sulfation pattern. Expression of enzymes involved in the Disclosure: No significant relationships. regulation of this sulfation pattern was highly affected by syndecan-1 over

iMig2012.org • Abstract Book 152 Poster Session 5 Epidemiology and Asbestos September 14, 2012 11:30-12:30

POSTER SESSION 5 September 14, 2012 11:30-12:30 POSTER SESSION 5 September 14, 2012 11:30-12:30

P5.20: NATIONAL SURVEY OF MALIGNANT MESOTHELIOMA AND P5.21: A GEOLOGICAL REVIEW OF CALIDRIA ASBESTOS AND ITS ASBESTOS EXPOSURE IN JAPAN AMPHIBOLE ASSEMBLAGE

1 2 3 4 Kenichi Gemba , Nobukazu Fujimoto , Katsuya Kato , Keisuke Aoe , Steven Kazan, William F. Ruiz poster sessions | september 14 Yukio Takeshima5, Kouki Inai5, Takumi Kishimoto2 Kazan Mcclain Lyons Greenwood & Harley, Oakland/CA/UNITED STATES OF 1Respiratory Medicine, Fukuyama Medical Center, Fukuyama/ AMERICA JAPAN, 2Respiratory Medicine, Okayama Rosai Hospital, Okayama/ JAPAN, 3Radiology, Okayama University Hospital, Okayama/ Background: Calidria asbestos is often generically described as a pure JAPAN, 4Medical Oncology, Yamaguchi-Ube Medical Center, Ube/ chrysotile asbestos. The carcinogenicity of pure chrysotile asbestos is JAPAN, 5Pathology, Nstitute Of Biomedical & Health Sciences Hiroshima disputed by the Union Carbide Corporation, which mined and sold Calidria, University, Hiroshima/JAPAN as well as manufacturers who used Calidria as an ingredient in asbestos containing products. The “chrysotile is safe” theory remains contrary Background: A newspaper article published in June 2005 reported that to medical and scientific evidence and has not been adopted by any five residents who lived near the now-closed asbestos cement pipe plant regulatory agency. The fact that chrysotile asbestos is carcinogenic is well in Amagasaki, Japan, developed pleural mesothelioma. The asbestos- published and is not the focus of this article. Rather this article challenges related problems that the article described caused considerable social the premise that all Calidria was pure chrysotile. Of the many grades of concern, resulting in the so-called “Kubota shock”. Asbestos has attracted Calidria, only one type was subject to a purification process designed to increasing social attention, but no large-scale studies have been conducted yield pure chrysotile asbestos. Manufacturing documents describe the to date investigating the clinical features of Malignant Mesothelioma (MM) other grades of Calidria as 90% chrysotile fiber with the remaining 10% in Japan. being naturally occurring impurities. This article focuses on the naturally occurring amphiboles found in the New Idria district where Calidria was Methods: In the present study, MM cases in Japan were investigated mined. retrospectively. We extracted records for 6030 cases of death due to MM between 2003 and 2008 to clarify the clinical features of MM, including its Methods: Geologic provenance, reported metamorphic mineral association with asbestos exposure (AE). assemblages, and watershed erosion principals are reviewed, including data from the Environmental Protection Agency, Benitoite gem Results: Of all these cases, a clinical diagnosis of MM was confirmed prospecting, and United States Geological Survey topography mapping. for 929. The origin of MM included the pleura in 794 cases (85.5%), the Fibrous amphibole found in blueschist shear zones are considered in three peritoneum in 123 cases (13.2%), the pericardium in seven cases (0.8%), dimensional space in order to understand the effect of rain and wind and the testicular tunica vaginalis in five cases (0.5%). The histological translocation and the significance of Franciscan rock inclusions caused by international mesothelioma interest group subtypes of MM included 396 epithelioid (55.9%), 154 sarcomatoid San Andreas Fault tectonics. (21.7%), 126 biphasic (17.8%), and 33 cases (4.7%) classified as “other types”. Of all the MM cases, AE was indicated in 76.8% and pleural Results: Fibrous amphibole inclusions, primarily tremolite and actinolite, plaques were detected in 34.2%. The number of asbestos particles was are found throughout the New Idria district and former Calidria mine in determined in 103 cases of MM. More than 1000 asbestos particles per a homogenous pattern due to the region’s geology and commingling of gram dried lung tissue were detected in 74.8% of cases and more than minerals from erosion, wind and rain. This mineral distribution is further 5000 particles were detected in 43.7% of cases. We compared patient supported by findings of fibrous amphibole in Benitoite gem claims characteristics and the diagnostic procedures for MM before and after the immediately adjacent and above the Calidria mine site. “Kubota shock”. Compared with the early phase of this study (2003–2005), the median age at diagnosis of MM was higher, the number of cases without definite diagnosis of MM was lower, the proportion of cases diagnosed by thoracoscopy was higher, and the percentage of cases in which the occupational history was described in the medical records was significantly higher in the later phase (2006–2008).

Conclusion: Our study confirmed that more than 70% of MM cases in Japan are associated with AE. The “Kubota shock” may affect some features pertaining to MM.

Disclosure: No significant relationships.

Conclusion: [Not applicable for this presentation.]

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 153 POSTER SESSION 5 September 14, 2012 11:30-12:30

P5.22: CASE REPORT: DEMONSTRATING THE INACCURACY OF DEATH CERTIFICATES

Steven Kazan Kazan Mcclain Lyons Greenwood & Harley, Oakland/CA/UNITED STATES OF AMERICA

Background: It has long been established1 that the incidence of mesothelioma is underreported on death certificates, leading to an underestimation of risk from asbestos exposure.2 We present a classic illustration demonstrating that this problem still exists today. Mike Hoffhine was born in 1943 and spent six years as a naval electrician, first on active duty on U.S. Navy ships, and then as a civilian employee at Hunters Point Naval Shipyard in San Francisco. In April 2001, an open right pleural biopsy demonstrated malignant epithelioid mesothelioma and pleural plaque. In June 2001, he underwent a resection of right middle and upper poster sessions | september 14 lobe segments, partial excision of his diaphragm, and partial excision of right parietal and visceral pleura by Dr. David Jablons at the University of California, San Francisco, with pathologic confirmation of the diagnosis and a AJCC/UICC classification pT2NoMx.

Methods: In August 2001, suit was filed against a group of defendants

Conclusion: This is a classic asbestos-related mesothelioma death. It international mesothelioma interest group demonstrates that even today, physicians misreport causes of death on death certificates, confirming yet again the inaccuracy of epidemiologic studies based on death certificate causes of death. 1Connelly, et al. Demographic Patterns for Mesothelioma in the United States. Journal of the American Cancer Institute, Vol. 78, No. 6, p.1058, June 1987. 2Kopylev, et al. Monte Carlo Analysis of Impact of Underascertainment of Mesothelioma Cases on Underestimation of Risk. The Open Epidemiology Journal, Vol. 4, 2011.

Disclosure: No significant relationships.

POSTER SESSION 5 September 14, 2012 11:30-12:30

P5.23: PATIENTS’ RIGHTS TO COMPENSATION FROM UNITED STATES BANKRUPTCY TRUST FUNDS and resolved after defendants thoroughly investigated Mike’s medical and Steven Kazan asbestos exposure history. Kazan Mcclain Lyons Greenwood & Harley, Oakland/CA/UNITED STATES OF AMERICA Results: Mike Hoffhine died in Missouri in April 2012. His death certificate listed the cause of death as respiratory failure due to “metastatic Background: Approximately 70 United States companies have filed for pulmonary malignancy,” said it was unknown whether tobacco use bankruptcy reorganization in whole or in part as a result of asbestos contributed to the death, and recorded a natural rather than accidental or liabilities. Approximately 40 of them have emerged from bankruptcy with industrial death. established trust funds which have already paid out about $10 billion, with another $30 billion remaining on hand. In addition, another ten or so bankruptcy reorganizations are pending and should emerge with $10-$15 billion in new Trust Funds within the next several years. American asbestos victims have full access to our court system and can recover from

iMig2012.org • Abstract Book 154 remaining corporate entities as well as Trust Funds, when appropriate. POSTER SESSION 5 September 14, 2012 11:30-12:30 All U.S. Bankruptcy Trust Funds are charged with responsibility to pay claims against the establishing company whether or not those claims P5.25: HEALTH EFFECTS AND RISK ASSESSMENT AMONG arose in the United States. As a result, foreign patients who have never RESIDENT POPULATION WITH NATURALLY OCCURING ASBESTOS been exposed to asbestos in the United States may still be entitled to EXPOSURE IN SOUTH OF ITALY present claims against these U.S. Trust Funds. A successful claim requires: (1) a mesothelioma diagnosis; (2) exposure to products for which that Tommaso Massaro1, Antonio Baldassarre1, Silvano Dragonieri2, Saverio Trust bears responsibility; (3) existence of a claim under local law; and Fiore3, Antonio Lettino3, Gabriella Cauzillo4, Angelo Caputo5, Filippo (4) determination of value under local law. This presentation will review Cassano1, Marina Musti1 the identity of existing and expected Trust Funds, the magnitude of their 1Dim - Occupational Medicine, University Of Bari, Bari/ITALY, 2Department individual assets, how each individual Trust approaches the value of Of Respiratory Diseases, University Of Bary, Bari/ITALY, 3Institute Of mesothelioma claims under American law, and ways in which exposure Methodologies For Environmental Analysis, National Research Council, Tito to Trust asbestos-containing products can be established. This includes a Scalo/ITALY, 4Cor Basilicata, Potenza/ITALY, 5Asp, Lagonegro/ITALY presentation concerning established and recognized non-U.S. work sites (in excess of 3,000 sites) and approved ships (in excess of 13,000 specific Background: Asbestos is contained in rock and soil in a Basilicata’s rural identified ships). We will briefly review the specific steps required to areas, in South of Italy. The characterization was achieved by mineralogical establish a claim and discuss the role of diagnosing and treating physicians and geological surveys. There is Tremolite in outcrops of serpentinite, in assisting patients to present such claims. amphibolite, metabasites, as well as in soils, bodies of landslides poster sessions | september 14 and debris. Environmental monitoring showed near the population Methods: Not applicable to this presentation. centers doses of Tremolite airborne between 1ff/L and 5ff/L. After the identification of cases of mesothelioma arisen in such areas as a result Results: Not applicable to this presentation. of exposure to natural sources of asbestos, a Consensus Conference was convocated to draw the guidelines for Health Surveillance, for preventive Conclusion: Not applicable to this presentation. measures and communication of the risk. The aim of this study was to evaluate the prevalence of asbestos-related diseases in people exposed Disclosure: No significant relationships. to tremolite and contribute to the determination of the risk assessment of exposure.

Methods: Epidemiological and health surveillance was carried out to POSTER SESSION 5 September 14, 2012 11:30-12:30 assess the prevalence of respiratory diseases associated with occupational and environmental asbestos exposure based on voluntary recruitment P5.24: DISEASE BURDEN OF MALIGNANT PLEURAL of population at risk among the residents. People underwent a health MESOTHELIOMA: TAIWAN EXPERIENCES plan that includes standardized questionnaire, occupational anamnesis,

1 1 2 pathological anamnesis, physical examination, counselling, spirometry Lukas J. Lee , Yu-yin Chang , Jung-der Wang (ATS criteria), pulmonary examination, chest radiographs examined by 1Division Of Environmental Health And Occupational Medicine, National 2 two B-readers. Selected cases could be subjected to HRCT. The risk Health Research Institutes, Zhunan, Miaoli County/TAIWAN, College Of assessment of exposure to Tremolite was performed with environmental Medicine And Hospital, National Cheng Kung University, Tainan/TAIWAN and personal sampling. The analysis of the filters was done with SEM and the characterization of the fibers with EDS method. Background: The burden of disease caused by asbestos have a substantial impact on quality and quantity of life in general population with potential Results: 699 people underwent the surveillance (332 men and 367 exposure from the workplace and residential environment, as well as may women). 7.1% of those examined have pleural plaques (10.8% of men, exert financial impact on patients, their families and the society. However, 3,8% of women). Interstitial Pneumopathies prevalence is 4%, (6,3% of the economic costs of asbestos-related diseases to the society remain to men, 1,9 % of women). Two cases of mesothelioma and one case of lung international mesothelioma interest group be elucidated. It is important to quantify the disease burden caused by cancer were found. 93% of asbestos-related diseases concerns farmers and malignant pleural mesothelioma (MPM) to facilitate the outcome research builders. Environmental monitoring of asbestos widespread showed that of and cost-effectiveness analysis for prevention of asbestos-related diseases. 368 samples in 8.9% of cases there was an excess of 2 ff/l with peaks up to 31.7 ff/l at construction sites for the safety of roads built on contaminated Methods: We retrieved cases of pleural cancer registered in the rocks. Personal sampling have involved 30 residents employed in different catastrophic illnesses of the National Health Insurance Research work activities: 5 builders , 15 farmers, 10 employees in jobs that do not Database (NHIRD) to estimate the survival functions for malignant pleural involve contact with soil. An exposure exceeding 2 ff/l occurred in 60% of mesothelioma, an asbestos-related malignancy. We also retrieved data farmers with a peak of 23.06 ff/l, in 100% of builders until 12.02 ff/l and in from the Taiwan Cancer Registry (TCR) and linked them with the National any case of employees in activities that not involving contact with soil. Mortality Registry to cross-validate the results. Assuming a constant excess hazard, we extrapolated lifetime survival function by the Monte Conclusion: Health surveillance has demonstrated the presence of Carlo method. For each MPM patient, we simulated an age- and gender- asbestos-related diseases both benign and malignant in residents in areas matched person without cancer based on the vital statistics of Taiwan to contaminated with Tremolite. 2 cases of mesothelioma in male farmers estimate life expectancy and expected years of life lost (EYLL). By using the have been found in 699 people examined, with a crude rate of 3 cases reimbursement data from the NHIRD, we calculated the average monthly in 1000, while the Italian national average for males is 3.49 cases per healthcare expenditures, which were summed to estimate the lifetime 100000 (Marinaccio 2012). According to the findings of health surveillance, healthcare expenditures after adjusting for the corresponding monthly personal and environmental sampling have documented the existence of survival probability. a risk to the general population, however, contained within 2 ff/l, while people employed in building and agriculture are exposed to higher doses of Results: A total of 503 cases of MPM were identified in the NHIRD during Tremolite for actions of soil disturbance. Exposure to Tremolite in this area 1997 ~ 2008. The average EYLL was predicted to be 15.1 (95% confidence is an occupational hazard. interval: 14.1 ~ 16.1) years, and the lifetime healthcare expenditures with a 3% annual discount were predicted to be $19,615 (95% CI: 14,505.3 ~ Disclosure: No significant relationships. 24,724.7) US dollars.

Conclusion: The burden of MPM, in terms of EYLL and lifetime healthcare expenditures, was substantial. Such estimates may provide useful empirical evidence for clinical and health policy-making.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 155 POSTER SESSION 5 September 14, 2012 11:30-12:30 (LDCT) in 26 institutions in Japan. Among them, 6286 (64.1%) subjects underwent subsequent screening after 2 years of interval. Clinical P5.26: RISK OF MALIGNANT MESOTHELIOMA AFTER 40 YEARS information such as histories of residential and occupational asbestos SINCE FIRST EXPOSURE: A POOLED ANALYSIS. exposure for all life was reviewed. Images were interpreted independently by 15 experienced radiologists or pulmonologists. Alison Reid1, Nicholas De Klerk2, Geoffrey Berry3, Corrado Magnani4, Daniela Ferrante5, Arthur W. Musk6, Enzo Merler7 Results: Self-reported history of occupational exposure was definitely and 1School Of Population Health, University Of Western Australia, Crawley/ possibly present in 1103 (11.2%) and 1702 (17.3%) subjects, respectively, WA/AUSTRALIA, 2Institute For Child Health Research, University Of Western whereas self-reported history of residential exposure was definitely and Australia, Subiaco/WA/AUSTRALIA, 3School Of Public Health, University Of possibly present in 262 (2.7%) and 931 (9.5%) subjects, respectively, Sydney, Sydney/AUSTRALIA, 4Translational Medicine, University Of Eastern although asbestos factory in their residential areas actually existed in 2870 Piedmont, Piedmont/ITALY, 5Dept.Of Translational Medicine, University (29.3%) subjects. Pleural plaque, pleural thickening, and non-calcified Of Eastern Piedmont, Piedmont/ITALY, 6Respiratory Medicine, Sir Charles pulmonary nodule/mass were identified in 264 (2.7%), 245 (2.5%), and Gairdner Hospital, Nedlands/WA/AUSTRALIA, 7Occupational Health 1003 (10.2%) subjects, respectively, on LDCT, resulting in approximately 4 Department, Local Health Authority, Venetian Mesothelioma Register, times higher detectability compared with chest radiography. As malignant Padova/ITALY tumor, lung cancer was pathologically confirmed in 29 (0.3%) subjects, including 3 subjects with pleural plaque on LDCT. However, self-reported Background: The risk of malignant mesothelioma increases proportionatley history of asbestos exposure was not identified in them. Similarly, it poster sessions | september 14 to the cumulative asbestos exposure and to the 3rd or 4th power of time was not confirmed in 77 (29.2%) of 264 subjects with pleural plaque on since first exposure. Most epidemiological studies do not have follow up LDCT. Based on logistic regression analysis, presence of pleural plaque beyond 40 years. on LDCT was significantly correlated with male (odds ratio OR, 2.32), age 60 years and more (OR, 1.75), smoking (OR, 1.60), self-reported history Methods: The data from 6 cohorts of exposed workers (three Italian of asbestos exposure (OR, 3.92), residential period in asbestos factory railway workers’ cohorts, one Amosite workers’ cohort, the Eternit area (OR every 10 years, 1.13), and asbestos-related work period (7 works cohort, and the Wittenoom workers’ cohort) and two cohorts of people identified, OR every 10 years, 1.30-3.21). Similarly, presence of lung cancer with residential exposure (Wittenoom residents and Eternit wives) has was significantly correlated with age 60 years and more (OR, 2.67) and been pooled. A nested case control design matched cases and controls presence of pleural plaque (OR, 4.17). After repeated LDCT screening, on calendar period and age. Conditional logistic regression and fractional among 6286 subjects consisting of 139 and 6147 subjects with and without polynomials were used to model the relationship between time since first pleural plaque on LDCT at baseline screening, respectively, 5 (3.6%) of exposure and risk of mesothelioma. Further models were developed to 139 and 7 (0.1%) of 6147 subjects showed marked progression and new adjust for increasing mortality from competing other causes. onset of pleural plaque, respectively. Furthermore, lung cancer was pathologically confirmed in 8 (0.1%) subjects. Results: The combined data consisted of 22,048 people with asbestos exposure (16,279 males, 5,769 females). There were 649 cases of Conclusion: Our results indicate the potential risk of asbestos exposure confirmed pleural mesothelioma (494 males, 155 females) and 142 cases among Japanese general population and also the importance of public of peritoneal mesothelioma (112 males and 30 females). Median time since relations and enlightenment for asbestos exposure among them. first exposure was 38.2 years (IQR 26.5-46.6). Median duration of exposure was 2.61 years (IQR 0.5-15.0). The risk of pleural mesothelioma increased Disclosure: No significant relationships. until 40 years since first exposure. An apparent lessening in the rate of increase after 40 years was only partly explained by allowing for competing risks. The peritoneal mesothelioma rate continued to increase. POSTER SESSION 5 September 14, 2012 11:30-12:30 Conclusion: The data from this large pooled international cohort of asbestos-exposed people indicates that the rate of pleural malignant P5.29: THE PREDICTIVE VALUE OF X-RAYS IN IDENTIFYING international mesothelioma interest group mesothelioma increases steadily until 40 years after first exposure and then ERIONITE-RELATED PLEURO-PULMONARY CHANGES OVER A the rate of increase appears to decrease. However the rate of peritoneal 6-YEAR PERIOD AMONG ADULTS LIVING IN TUZKOY, TURKEY. mesothelioma appears to continue rising steadily beyond 40 years. Salih A. Emri1, Seha Akduman2 Disclosure: No significant relationships. 1Medical Faculty Depts. Of Chest Diseases, Hacettepe University, Ankara/ TURKEY, 2Department Of Chest Diseases, Hacettepe University, Ankara/ TURKEY

POSTER SESSION 5 September 14, 2012 11:30-12:30 Background: Due to environmental exposure to erionite, malignant pleural mesothelioma (MPM) is a common tumor in three villages in Central Turkey. P5.27: JAPANESE GENERAL SCREENING STUDY FOR ASBESTOS- Of these, Tuzkoy is the most well known and the largest village. Between RELATED DISEASES (JGSARD): RESULTS OF 2 YEAR FOLLOW UP 1980 and 1988, MPM accounted for more than one-fifth of the total deaths and approximately half of the cancer-related deaths in Tuzkoy. Kazuto Ashizawa1, Nobuhiko Seki2, Kenji Eguchi2, Takuhiro Yamaguchi3, Masahiro Kaneko4, Masahiko Kusumoto5 Methods: Chest X-rays of 231 adults 18 years of age older living in Tuzkoy 1Clinical Oncology Center, Nagasaki University Hospital, Nagasaki/ were taken in 2006. Mortality outcomes were investigated retrospectively JAPAN, 2Department Of Internal Medicine, Teikyo University School Of in 2012 using verbal autopsy methods. The X-rays were examined and Medicine/JAPAN, 3Tohoku University Hospital/JAPAN, 4National Cancer compared to mortality outcomes to determine their predictive value in Center Hospital/JAPAN, 5Department Of Radiology, National Cancer Center identifying malignant mesothelioma (MM) over the 6-year period. Hospital/JAPAN Results: One hundred and forty-nine (64%) X-rays were of female patients. Background: The number of patients with pleural mesothelioma and lung The mean and median ages of patients were 47 ± 15 years and 47 years, cancer associated with asbestos exposure has recently been increasing respectively (Range: 18-85 years). Of the 231 X-rays, 56 (24%) were in Japan. The aim of this study was to prospectively evaluate the actual abnormal: 14 (6%) suggested pleural calcification; 11 (5%) had diffuse situation of asbestos exposure and the prevalence of asbestos-related pleural thickening; 13 (5.7%) had loss of the costophrenic angels; 5 (2%) diseases in a group of Japanese general population. had apparent thickening in the minor fissure; and 1 (<1%) had pleural effusion. Four chest x-rays were unreadable. Over the 6-year period, 15 Methods: From 2006 to 2008, 9810 subjects (mean age, 57 years; 54% deaths were recorded, 9 (4 males and 5 females) of which were due to MM: male and 50% smokers) underwent chest radiography and low-dose CT five died from pleural and 4 from peritoneal mesothelioma. Mean ages of

iMig2012.org • Abstract Book 156 death for males was 61.5 ± 10 (Range: 52 to 72), and for females, 60±9 POSTER SESSION 5 September 14, 2012 11:30-12:30 (Range: 49 to 73 years). Of the 9 patients who died from MM, only one had an abnormal chest X-ray; this patient was in treatment but subsequently P5.33: THE FEMALE FACE OF BRITAIN’S ASBESTOS CATASTROPHE died within a year of the 2006 X-ray. Laurie Kazan-Allen Conclusion: Chest X-rays were not predictive of MM-related deaths over International Ban Asbestos Secretariat, Stanmore/UNITED KINGDOM the 6-year period among adults living in Tuzkoy. Erionite-related MM tends to progress quickly, which is likely why only one case was detected Background: Considering the colossal levels of asbestos exposure in the 2006 X-rays. Based upon these findings, we suggest low dose experienced by British workers, consumers, bystanders and community non- contrast CT screening of the thorax with simultaneous abdominal members during the 20th century, there can be no doubt that the death toll ultrasonographic examination to detect cases in the early stages, especially from asbestos-related diseases has been massive. It is unfortunately true, in high risk areas like Tuzkoy. however, that no one knows precisely how many lives have been lost due to the country’s love affair with asbestos; how many families have been Disclosure: No significant relationships. torn asunder by avoidable asbestos-related deaths or how many children’s lives have been decimated by the early loss of a parent or the trauma of a beloved grandparent’s premature death. For years, Britain has had the unwelcome distinction of being amongst the countries worst affected POSTER SESSION 5 September 14, 2012 11:30-12:30 by the asbestos cancer, mesothelioma. Historically, male mesothelioma poster sessions | september 14 deaths have dominated the statistics with, at times, six times as many male P5.30: MALIGNANT MESOTHELIOMA AND OCCUPATIONAL OR as female fatalities. Despite the lower death rate amongst British women, it BYSTANDER ASBESTOS EXPOSURE: A STUDY OF 384 CASES is of interest to note that so many of the landmark cases through which the national asbestos reality has been revealed relate to the personal tragedies Jennifer Faig, Edward Levine, Jill Ohar of female victims. This paper will examine the asbestos experiences of Wake Forest University School Of Medicine, Winston-salem/NC/UNITED seven British women against the backdrop of the emerging asbestos STATES OF AMERICA catastrophe. Those whose fates were sealed by fatal exposures to asbestos covered a wide spectrum of society from factory workers to schoolteachers Background: Exposure to asbestos fibers in either an occupational or to civil servants; environmental and para-occupational asbestos exposures non-occupational setting is associated with the development of malignant transformed what was formerly regarded as an occupational epidemic into mesothelioma. The purpose of this study is to analyze 384 mesothelioma a public health crisis. patients with documented occupational or bystander asbestos exposure and to report the anatomical location and survival outcomes of patients in Methods: The research for this paper has included a study of published reference to exposure type. papers, archival material and interviews.

Methods: The 384 subjects selected for this study had known exposure Results: The findings of this paper confirm the transformation of what was to asbestos and a diagnosis of mesothelioma. The cohort of subjects initially believed to be an occupational health issue into a public health consisted of patients referred to Wake Forest University Baptist Medical crisis. Recognizing the asbestos hazard, Britain, like other industrialized Center for surgical management, attendees of the Mesothelioma countries, banned the import, use and sale of asbestos. Nevertheless, 2 Applied Research Foundation (MARF) conference, and law firm subjects million tonnes are still being used every year in the developing world. The undergoing legal action for occupational exposure. Data was obtained from British experience suggests that calls for a global ban on asbestos are both questionnaires and medical records. justified and urgent.

Results: There were 286 patients exposed to asbestos in an occupational Conclusion: The profits of Britain’s asbestos sector were built on the setting: 259 (90.6%) developed pleural mesothelioma, 27 (9.4%) deaths of these and other asbestos victims. Corporate executives as developed peritoneal mesothelioma. Mean age at first exposure was well as government ministers, civil servants and elected representatives international mesothelioma interest group 18.3 years (SD 9.4). Mean age at diagnosis was 67.3 years (SD 11.2). Mean were responsible for unleashing a ferocious onslaught on ordinary men latency was 49.1 years (SD 10.9). Median survival was 19.7 months. There and women who were powerless in the face of this deadly carcinogen. were 98 patients exposed to asbestos in a non-occupational setting: 46 The same excuses advanced to prolong the use of asbestos in Britain (46.9%) developed pleural mesothelioma, 52 (53.1%) developed peritoneal are still being promoted by vested interests in countries where asbestos mesothelioma. Mean age at first exposure was 9.1 years (SD 10.2). Mean use remains legal. The dimensions and severity of the British asbestos age at diagnosis was 51.3 years (SD 16.3). Mean latency was 42.7 years experience should be more than enough to convince a reasonable person (SD 15.5). Median survival was 56.7 months. There was no significant that humanity has a right to live in an asbestos-free atmosphere difference in survival between bystanders and primary exposure after adjusting for age at diagnosis and gender (p=0.86). Irrespective of exposure Disclosure: No significant relationships. status, peritoneal mesothelioma patients had significantly better survival (HR=0.36 p <0.001) compared to thoracic mesothelioma patients, after adjusting for age at diagnosis and gender.

Conclusion: The observed findings suggest that individuals with occupational asbestos exposure are more likely to develop pleural than peritoneal mesothelioma whereas bystanders develop either pleural or peritoneal mesothelioma. Although there was no survival difference between the exposure groups, peritoneal patients had an overall better survival than pleural patients. This study emphasizes the importance of assessing the patient’s environmental exposure history. Early recognition of this risk factor can hopefully alter the prognosis of this dreadful disease.

Disclosure: No significant relationships.

iMig2012.org • Abstract Book 157 Poster Session 5 Peritoneal Mesothelioma September 14, 2012 11:30-12:30

POSTER SESSION 5 SEptember 14, 2012 11:30-12:30 And Oncologic Surgery, University Of Maryland School Of Medicine/UNITED STATES OF AMERICA P5.34: THE ROLE OF PERIOPERATIVE SYSTEMIC CHEMOTHERAPY IN DIFFUSE MALIGNANT PERITONEAL MESOTHELIOMA PATIENTS Background: Malignant peritoneal mesothelioma (MPeM) is a cancer of TREATED WITH CYTOREDUCTIVE SURGERY AND HYPERTHERMIC the abdomen that frequently recurs after surgical resection and is relatively

resistant to chemotherapy. We have recently reported that over expression poster sessions | september 14 INTRAPERITONEAL CHEMOTHERAPY of PI3K and mTOR pathways in tumor samples from patients with MPeM are associated with shortened patient survival. In this study our aim was to Marcello Deraco1, Dario Baratti1, Rossella Bertulli2, Shigeki Kusamura1, test the effectiveness of a novel class I PI3-Kinase specific inhibitor for the Ionut Hutanu3 treatment of MPeM. 1Surgery, Fondazione Irccs Istituto Nazionale Tumori, Milano/ITALY, 2Adult Mesenchymal Tumor Medical Oncology Unit, Fondazione Irccs Istituto Methods: MPeM cells (Meso-1, Meso-2 and Meso-3) were treated with Nazionale Tumori, Milano/ITALY, 3Surgical Department, University Of BKM120 (50 nM/L) and cell proliferation rates were assessed in vitro. Medicine And Pharmacy ‘Gr T Popa’, Iasi/ROMANIA Downstream PI3K/mTOR signaling was studied by analyzing RNA by qPCR and protein expression by Western blot. Anti-tumor effects of BKM120 were Background: Provided that studies on CRS and HIPEC to treat DMPM tested in an MPeM murine xenograft model. usually have not explored the prognostic significance of perioperative systemic treatments, we attempted to evaluate the effects of preoperative Results: BKM120 treatment for 48 h significantly (p<0.05) inhibited MPeM systemic chemotherapy (CT) on overall (OS) and progression free survivals cell proliferation. There was no clear difference in anti-proliferative activity (PFS). As secondary endpoint we evaluated the effects of preoperative of BKM120 in phosphatase and tensin homolog (PTEN) positive versus PTEN systemic chemotherapy on short-term surgical outcomes regarding null cell lines. Treatment of cells with BKM120 significantly reduced PI3K morbidity and completeness of cytoreduction. downstream signaling; PI3K-p110α, phospho p85, AKT and phospho AKT protein expression were markedly less in treated cells compared with the Methods: Data from 116 DMPM patients treated with CRS and HIPEC control group. Although BKM120 is a specific inhibitor of class I PI3-kinase, from August 1995 to October 2011 were retrospectively analyzed from a mTOR and phospho mTOR expression were also reduced in treated cells. prospectively collected database in the NCI of Milan. Sixty cases underwent S6 kinase activity showed no significant difference in the control versus preoperative CT, 30 underwent postoperative CT, and 26 did not perform treated groups whereas phospho S6 kinase activity decreased in BKM120 any CT. Fifty-five cases used the perioperative combination of platinum treated group. Oral treatment of BKM120 (20mg/every alternate day) for and pemetrexed. We tested whether covariates related to clinical, one month significantly reduced xenograft growth compared to placebo histologic, perioperative CT, and surgical treatment were correlated with controls. completeness of cytoreduction (CC), postoperative G3-5 morbidity, and progression free (PFS) and overall survivals (OS). Univariate and multivariate

Conclusion: BKM120 is a potential inhibitor of PI3K and mTOR signaling in international mesothelioma interest group analysis were performed. MPeM. Tumor growth can be suppressed in MPeM xenograft models with oral administration of BKM120. Further studies are warranted to evaluate Results: factors independently associated with CC were ECOG the action this compound in tumor relapse and PI3K associated pathways. performance status (PF) =0, and PCI <20. Factors independently associated This work was supported by a grant from the Mesothelioma Applied with postoperative G3-5 morbidity were ECOG PS>1, bowel anastomosis Research Foundation (MARF) and number of peritonectomy procedures. Preoperative platelet count>400x103/mm3, histological subtype (biphasic and sarcomatoid vs Disclosure: No significant relationships. epithelial), CC, and G3-5 morbidity were independent prognostic factors. Preoperative CT was not associated with CC or G3-5 morbidity. There was no significant difference in terms of survival between the preoperative CT, postoperative CT and no-CT groups. POSTER SESSION 5 September 14, 2012 11:30-12:30 Conclusions: the CC, G3-5 and OS were not influenced by aspects related P5.36: MESOTHELIOMA CLUSTER IN A MEDIUM SIZED HOSPITAL to peritoperative CT. Such a comparative analysis should be repeated in a multi-institutional context with a judicious selection of control groups well- IN SOUTH-EAST CHINA: RELATIONS TO ASBESTOS EXPOSURE balanced regarding the distribution of risk/prognostic factors. AND CLINICOPATHOLOGICAL FINDINGS

1 2 Disclosure: No significant relationships. Gao Zhibin , Oluf D. Røe 1Department Of Pathology, Yuyao Peoples Hospital, Yuyao, Zhejiang Province, /CHINA, 2Department Of Cancer Research And Molecular Medicine, Norwegian University Of Science And Technology (Ntnu), POSTER SESSION 5 September 14, 2012 11:30-12:30 Trondheim/NORWAY

P5.35: AN ORAL PAN-CLASS I PI3-KINASE INHIBITOR, NVP- Background: Asbestos, including chrysotile, is a Group I carcinogen BKM120, INHIBITS TUMOR GROWTH IN HUMAN MALIGNANT according to the IARC. In China chrysotile asbestos is still produced and consumed but there are few reports that link it to mesothelioma incidence. PERITONEAL MESOTHELIOMA MURINE XENOGRAFTS We here present a cluster of malignant mesothelioma related to chrysotile asbestos exposure diagnosed in a medium-sized hospital of Yuyao in Sheelu Varghese1, H Richard Alexander2 the Zhejiang Province of South-East China and their clinicopathological 1Division Of General And Oncologic Surgery, University Of Maryland School characteristics. Of Medicine, Baltimore/UNITED STATES OF AMERICA, 2Division Of General

iMig2012.org • Abstract Book 158 Methods: Consecutive patients registered at the pathology department at the Yuyao Peoples Hospital with suspected mesothelioma diagnosis by clinical findings and histology by Hematoxylin-Eosin (HE) staining from 2005- 2010. The histological diagnosis was verified through immunohistochemistry with the positive markers calretinin, WT1, D2-40, CK5-6, AE1-AE3, CAM5.2, EMA and the negative markers CEA, BerEP4, MOC31, TTF-1 and the hormone receptors ER and PgR. Asbestos exposure data were obtained from the patient files and and/or by contacting the patients relatives.

Results: Among 38 suspected mesothelioma cases based on HE and clinical picture, 27 were verified with immunohistochemistry, 16 peritoneal and 11 pleural. Peritoneal mesothelioma (n=16) were all females with a mean age at diagnosis of 51 years (range 43-71) and of those were occupational information could be obtained (n=13) there was a 92% occupational asbestos exposure by hand spinning of asbestos ranging from 1-12 years. The females with pleural mesothelioma (n=10) had a mean age at diagnosis of 57 years (range 38-82) and an occupational poster sessions | september 14 asbestos exposure by hand spinning of asbestos of 100% ranging from 7-10 years. Only one male aged 67 was registered with pleural mesothelioma, with unknown asbestos history. Median survival was only 5 months on the cases where survival was registered (n=7). Revised diagnosis after immunohistochemistry in 11 cases included various carcinomas, among them a mucinous adenocarcinoma, uterine cervix carcinoma and pulmonary adenocarcinoma. 4/9 females (44%) had a history of heavy asbestos exposure, ranging from 5-20 years of hand spinning of asbestos. Three males also had a revised diagnosis, two with carcinoma where one had worked 5 years in an asbestos factory, and one benign asbestosis.

Conclusion: In this retrospective study of a mesothelioma cases from a single hospital in Yuyao, there were gross overrepresentation of women, 38:1, and of peritoneal mesothelioma, 68%. Importantly, 90-100% of these cases, where an occupational history was obtained, had a history of several years of asbestos by hand spinning. In a study published in 1985 from Cixi, a city close to Yuyao, described small asbestos factories and household spinning with high registered asbestos dust concentration were women used to work from10 to 12 hours daily [1]. The median latency time of mesothelioma from asbestos exposure is 40 years, so the women described here were likely exposed very young as the mean age at diagnosis was only 51. Moreover, other cancers were seen in these highly exposed individuals. As the Yuyao area still harbor small asbestos factories, such clusters of mesothelioma will be expected in the future.

Disclosure: No significant relationships. international mesothelioma interest group

iMig2012.org • Abstract Book 159 Author Index Presentation numbers in bold represent the presenting author.

A Baratti Dario...... P5.34, VA1.2 ...... P2.14, P4.10, WSXI.5 Chaouche-Mazouni Siham...... P5.15

Barbieri Stefano...... IIIA.5 Boyer Michael...... IVB.4 Cheah Huimin...... IVA.3 index author Abdalsamad Issam...... IVC.6 Barbone Dario...... IIIB.2, IIIB.3 Bracci Elisa...... P4.22 Chellini Elisabetta...... IIIC.3 Abtin Fereidoun...... IIB.3, IIB.6 Barlow Juliann...... IIB.4, IVC.5 Brambilla Elisabeth...... IVC.6 Chen Aileen B...... IIIA.2, P1.08 Adachi Shuichi...... P1.34 Barr Martin P...... IIA.7 Braun Julie...... P4.33 Chen Jin Li...... VA1.4 Aebersold Ruedi...... P3.04 Barrington Sally...... P2.20 Braune Nico...... P4.17 Chen Katherine...... VA1.4 Aerts Joachim G...... IIC.2, IIC.7, Batirel Hasan F...... P1.03, P3.13, P4.12 Bressan Vittoria...... IIIC.5 Chen Lu...... P4.11 ...... P1.30, P1.32, P3.26 Battula Sailaja...... IIIB.2, IIIB.3 Brighton David...... P2.31 Chen Ya-Fang...... P2.27 Agarwal Vijay...... P5.01 Baughn M...... P4.02 Broaddus Courtney...... IIIB.2, IIIB.3 Chen Zhuo...... P4.11 Ahiskali Rengin...... P1.03 Bauman Julie...... IB.3 Brochard Patrick...... IVC.6 Cheng Ngan C...... IC.5 Ak Guntulu...... P3.13 Bausch-Fluck Damaris...... P3.04 Brody Edward...... IA.2 Cheng Tiffany...... IVA.2 Akduman Seha...... P5.29 Bazhenova Lyudmila...... P4.02, IB.2 Bromage Hediye E...... P5.07 Cheng Yeun Yee...... IC.5, P4.24, Albelda Steven M...... IVB.3, Bech Cecilia...... IA.6 Brown Chris...... IVB.4 ...... WSXI.4, WSXI.5 ...... IVB.6, P1.06, P3.15 Becich Michael J...... P1.24 Brown Matthew D...... P2.15 Cheng Yuen Y...... P5.14 Alexander H. Richard...... P5.35 Begueret Hugues...... IVC.6 Buech Thomas...... P3.19 Cheng Yuen Yee...... IIA.2 Alexeyenko Andrey...... IC.6, P5.17 Bendell Johanna...... IVB.2 Bueno Raphael...... IA.7, IIB.4, IIIB.2, IIIB.3, Chernova Tatyana...... P5.05, P5.16, VA2.2 Allen Aaron M...... P3.27 Bendinelli Sara...... P4.22 ...... IVC.5, P1.08, IIIA.2, P1.21, P4.20, Chew Shan Hwu...... P5.06 Alley Evan...... IIIA.4, P1.19, P1.20, Bennett Jonathan...... P5.05, P5.16 ...... VB.5, VC.3, VIC1.2, VIC1.3, VIC2.1 Chicklore Sugama...... P2.20 ...... P2.01, P2.02, P3.15 Beretta Giordano D...... P4.27 Bunck Alexander C...... P2.22 Chirieac Lucian...... P1.25, P2.23, P2.24 Allo Ghassan...... P3.02 Berger Walter...... IVA.6, P4.07 Burris Howard...... IVB.2 Chirwa Tobias...... IIIC.4 Amatya Vishwa J...... P4.01 Berry Geoffrey...... P5.26 Busacca Sara...... IIIB.4, P4.25, P4.29 Cho John...... IIB.5, P3.10, P3.27 Ambrogi Vincenzo...... P3.23 Berthelsen Anne K...... P3.21 Bushell Martin...... P5.05, P5.16, VA2.2 Choi Meena...... P3.04 Amessis Malika...... P3.29 Bertulli Rossella...... P5.34, VA1.2 Butnor Kelly J...... VIA.3 Churcher Katheryn...... VIB1.1 Amin Waqas...... P1.24 Besse Benjamin...... IA.4, P1.13 Butte Atul...... WSXI.3 Ciminale Vincenzo...... P5.12 Amirkhani Ardeshir...... P3.08 Bethel Kelly...... P4.02 Buyske Jo...... P2.03 Clarke Belinda...... P1.35, P2.14, P4.10 Ampollini Luca...... IIIA.5 Betta Pier Giacomo...... P4.21 Bölükbas Servet...... IIB.2, IIIA.3, Clarke Stephen...... IA.5, P1.31, P3.08, P4.34 Andersen Claus B...... P3.01 ...... P3.03, P3.05, P3.06 Betti Marta...... P4.21 Cleaver Amanda...... P2.15, P3.24 Andersen Morten...... P3.01 Beuschel Stacie...... P5.08, IC.7, VIA.5 Coer Andrej...... P2.10 Andujar Pascal...... P5.10, VB.4 Bibby David...... IVB.4 Colby Aaron...... VIC2.4 Anraku Masaki...... IIB.5, P4.11 C Bidlingmaier Scott...... VIA.4 Colmou Karen C...... P3.29

Aoe Keisuke...... P4.01, P4.30, P5.20 international mesothelioma interest group Bijelic Lana...... VA1.3 Colombo Paolo...... IIIA.5 Aoyama Katsuhiko...... P4.18 Cabras Antonello D...... VA1.2 Bilgi Zeynep...... P4.12 Colson Yolonda L...... VIC2.4 Arbiser Jack...... P5.09 Cadby Gemma...... P4.21 Bille’ Andrea...... P1.23, P3.18 Connor Theresa...... P3.24 Ardissone Francesco...... P4.21 Cain Kelvin...... P5.16 Billiot Fanny...... IA.4 Cook Alistair M...... IB.4, IIC.4 Arif Qudsia...... IC.2 Cale Alex...... P1.01 Birnie Kimberly A...... P5.03, WSXI.2 Cook Gary...... P1.23, P2.20 Armato Samuel G...... P1.16, P1.17, VC.5 Cameron Robert B.....IIB.3, IIB.6, P1.09, P3.14 Bitanihirwe Byron...... P1.33, P3.02 Coolen Johan...... VC.6 Armstrong Nicola J...... WSXI.4 Campion Loic...... P3.29 Black Brad...... IA.2 Copin Marie Christine...... P5.15, VB.4, IVC.6 Arns Madleine...... P4.07 Cangir Ayten K...... VC.4 Blackler Elizabeth...... P2.30 Cornelissen Robin...... IIC.2, P1.30 Arslan Sertac...... P3.09 Canino Claudia...... P5.04 Blakely Collin...... IIIB.2 Coskinas Xanthi...... IVB.4 Asamura Hisao...... VC.4 Cantoni Anna Maria...... IIIA.5 Blaney Jaine...... P4.25 Costa Barbara...... P4.22 Asanuma Kozo...... P4.18 Cao Shousong...... VIA.7 Blanquart Christophe...... P4.06 Costa Chrisostome...... P4.28 Ashizawa Kazuto...... P5.27 Capron Frederique...... IVC.6 Boisgerault Nicolas...... P3.25 Courtney Deborah...... P4.10 Aspesi Anna...... P4.21 Caputo Angelo...... P5.25 Boisot Saskia...... P4.02 Coussens Lisa M...... IIIB.2 Astoul Philippe...... IB.5 Carbognani Paolo...... IIIA.5 Boittiaux Irving...... P4.33 Cowen Michael...... P1.01 Atay Scott M...... VIC2.1 Carbone Michele...... IA.2 Boldrini Laura...... P4.22 Craig Caraline M...... P2.30 Audigier-Valette Clarisse...... IB.5 Casadio Caterina...... P4.21 Bomalaski John...... IVB.5 Creaney Jenette...... IB.4, IC.4, IVB.4, IVC.4, Avril Norbert...... IVB.5 Casalone Elisabetta...... P4.21 Bonassi Stefano...... P4.21 ...... P2.25, P4.05, VB.2, WSXI.1, Ayers Deborah...... IA.2 Casbard Angela...... P4.29 ...... WSXI.2, WSXI.4 Bonnette Pierre...... P3.29 Case Bruce W...... VIB2.1 Creveuil Christian...... IB.5 Bonventre Joseph...... P3.20, P4.19 Cassano Filippo...... P5.25 Cufer Tanja...... IB.6 Booton Richard...... P1.26 B Cauzillo Gabriella...... IIIC.3, P5.25 Culligan Melissa J...... IIIA.4, P1.19, P1.20, Borczuk Alain...... VA1.4 Cawkwell Lynn...... P5.01 ...... P2.01, P2.02, P2.03, P3.15 Bordignon Claudio...... P1.10, P1.15, P2.16 Baas Paul...... IB.6, VIC2.2 Cedres Susana...... P4.03, P4.04 Cummins Michelle...... IVB.4 Borroni Ester...... P2.13 Badrian Bahareh...... P5.03, WSXI.2, WSXI.4 Cellerin Laurent...... P4.06 Cummins R...... P5.18 Borsczuck Alan...... IVC.2 Bahleda Rastislav...... P1.13 Cemazar Maja...... P2.10 Cunniff Brian...... P5.09 Bosl Justin A...... P1.29 Baird Anne-Marie...... IC.3, IIA.7 Cengel Keith...... IIIA.4, P1.19, P1.20, Currie Andrew J...... P2.15 Bosshard-Carter Leticia...... VIA.1 Baldassarre Antonio...... P5.25 ...... P2.01, P2.02, P2.03, P3.15 Bott Matthew...... IIA.3, IIA.4 Baldini Elizabeth H...... IIIA.2, IVC.5, Cerciello Ferdinando...... P3.04 Botta Mario...... P4.26 ...... P1.08, P1.21, VC.3 Ceresoli Giovanni L...... P4.26, P4.27 Bowman Rayleen V...... IIA.2, P1.35, Baltic Svetlana...... IVA.3 Chang Yu-Yin...... P5.24

iMig2012.org • Abstract Book 160 D Elshazley Momen H.K.A...... IIA.6 ...... P3.11, P3.12, P4.09, P4.16 Harbut Michael...... IA.2, IA.3, P1.22 Emri Salih A...... P5.29 Fusari Antonella...... IIIA.5 Hardy James...... P2.04, P2.05 D’Alò Daniela...... IIIC.3 Endress Anthony...... P1.22 Fushimi Takuro...... P4.13 Harvey B.J...... P5.18 Daga Antonio...... P1.12, P2.12 Endress Carmen...... P1.22 Harvie Rozelle...... P1.31, P3.08 Danel Claire...... IVC.6 Ermerak Onur...... P1.03, P3.13, P4.12 Hasan Baktiar...... IB.1, IB.6 Dang Nam H...... P4.01 G Hasani Arman...... IB.4 Danson Sarah...... IB.6 Hase Tetsunari...... IIA.6 Dare Hanne...... P2.25 F Gaafar Rabab M...... IB.1, IB.6 Hasegawa Seiki...... P1.05, P2.08, P2.29, Darlison Liz...... P2.31 Galanis Evanthia...... IVB.7 �� P3.11, P4.16 Dasilva Marcelo...... IIB.4, P1.08, P1.21 Fabiny Robert...... VC.7 Galateau-Sallé Françoise...... IVC.2, IVC.6, Hasegawa Yoshinori...... IIA.6 De Bono Johan...... IVB.2 Faig Jennifer...... P5.30 ...... P1.27, P5.10, VB.4 Hashimoto Masaki...... P1.05, P2.08, P3.11 De Keyzer Frederik...... VC.6 Farace Francoise...... IA.4 Garbe Loulette...... IVC.67 Hashimoto-Tamaoki Tomoko...... P1.11, P3.07

De Klerk Nicholas...... IIIC.2, P5.26 Farberov Maria...... P2.30 Gattani Sumedha...... IC.5 Hasina Rifat...... IC.2 index author De Lajartre Anne Yvonne...... IVC.6 Fatato P...... IB.2 Gemba Kenichi...... P4.30, P5.20 Hassan Raffit...... IB.2, VIA.3 De Leyn Paul...... P1.18, VC.6 Favaretto Adolfo...... P4.26, P5.12 Gemignani Federica...... P4.22 Hatz Rudolf A...... P3.16, P3.17, P3.19 De Perrot Marc...... IIB.5, IIC.3, IIC.6, P1.36, Favoni Roberto E...... P4.22 Gennaro Valerio...... IIIC.3 Haustermans Karin...... P1.18 ...... P3.02, P3.10, P4.11, P4.15 Feigen Malcolm...... P3.27, VC.7, VIB1.1 Ghanim Bahil...... IVA.6, P4.07 Hauw Philippe...... P5.15 De Reynies Aurélien...... VB.4 Fein Luis...... IB.3 Gibbs Allen...... IVC.2 He Biao...... IVA.2 De Rienzo Assunta...... IA.7, P4.20, VB.5, Feld Ronald...... IB.3, IIB.5, P3.10 Gibney Barry C...... VIC2.4 He Xiaomin...... IB.3 ...... VIC1.2, VIC1.3, VIC2.1 Feldman Michael...... P1.24 Gilg Soit Ilg Anabelle...... IVC.6 Hebbel Robert P...... IVB.7 De Vincenzo Fabio...... P4.26 Felip Enriqueta...... P4.03, P4.04 Gill Ritu R...... IIIA.2, IVC.5, P1.08, Hegedus Balazs...... IVA.6, P4.07 De Wever Walter...... VC.6 Felley-Bosco Emanuela..... IIA.5, P1.33, P3.04 ...... P1.21, VC.3 Hegmans Joost P.J.J...... IIC.2, IIC.7, De Witte Evelien...... P1.18 Fennell Dean...... IC.3, IIIB.3, IIIB.4, Gilligan David...... IVB.5 ...... P1.30, P1.32, P3.26 De Zotti Renata...... IIIC.3 ...... IVB.5, P1.12, P1.26, P4.25, P4.29 Gilmore Denis M...... VIC2.4 Heidari-Hamedani Ghazal...... IC.6, P5.17 Decramer Marc...... P1.18 Fernandez-Madrid Felix...... IA.3 Ginsberg Michelle S...... IB.7 Heintz Nicholas...... P5.09 Degiovanni Daniela...... P4.26 Ferolla Elena...... P5.17 Giordano Laura...... P4.26 Heitz Markus...... IIIC.4 Dekkers Jacqueline...... IIC.7 Ferrante Daniela...... P4.21, P5.26 Giordano Mara...... P4.21 Held Paul...... P5.09 Delvenne Philippe...... P4.28, P4.33 Feske Sandra...... P3.16, P3.17 Girardi Paolo...... IIIC.5 Helou Joelle...... P3.29 Demelker Yvonne...... P2.25, WSXI.1 Fetsch Patricia...... VIC2.1 Giraud Philippe...... P3.27, P3.29 Henderson Douglas...... IA.5, P5.14 Demes Melanie...... P3.03, P3.06 Filipits Martin...... IVA.6, P4.07 Giroux Dori...... VC.4 Hikosaka Yu...... P2.09 Deraco Marcello...... P5.34, VA1.2 Fiore Saverio...... P5.25 Gledhill David...... P2.31 Hillegass Jedd M...... VIA.3, VIA.5 Descatha Alexis...... P5.10 Fiorito Giovanni...... P4.21 Glineur Corine...... P5.15 Hino Okio...... P1.34 Dhanasingh Immanuel...... P2.11 Fishbein Michael...... IIB.6, P3.14 Goldberg Eugene P...... P5.11 Hiraki Akio...... P4.01 Dhir Rajiv...... P1.24 Fisher Scott...... P3.24 Gonzalez Cindy...... P2.21 Hirata Tomomi...... P2.26 Di Gaetano Cornelia...... P4.21 Fisseler-Eckhoff Annette...... IIB.2, P3.03, Goodman Brian M...... IIIA.2, IVC.5 Hirayama Noriko...... P4.09, P4.16 Dianzani Irma...... P4.21 ...... P3.05, P3.06 Goparaju Chandra...... IA.2, P5.04 Hiroshima Kenzo..... IIIB.5, IVC.2, IVC.3, VIA.2 Dick Ian...... P2.25, P4.05, VB.2, WSXI.1 Flatberg Arnar...... P4.23 Goto Masashi...... P4.18 Hirota Tomomi...... P2.28 Dick Ian M...... P4.08 Flores Raja...... P1.24 Gounant Valerie...... IB.5 Hjerpe Anders...... IC.6, P1.28, Diez Claudius...... P4.17 Flynn Louise...... IIA.7 Grauslund Morten...... P3.01 ...... P3.09, P4.32, P5.17 Dipietrantonj Carlo...... P4.26 Fong Kwun M...... IIA.2, P1.35, P2.14, P4.10 Gray Steven...... IC.3, IIA.7, P1.12 Hmeljak Julija...... P2.10 Do Hanh...... IVA.2 Fontanini Gabriella...... P4.22 Gregoire Marc...... P3.25, P4.06 Hoang Chuong D...... WSXI.3

Dobra Katalin...... IC.6, P1.28, Fonteneau Jean-François...... P3.25 Gregorc Vanesa...... P1.10, P2.16 Hoda Mir A...... IVA.6, P4.07 international mesothelioma interest group ...... P3.09, P4.32, P5.17 Forastiere Francesco...... IIIC.3 Greillier Laurent...... IB.5 Hoffman Hans...... VC.4 Dolly Saorise...... IVB.2 Ford Victoria...... IIB.5 Griffiths Gareth...... P4.29 Hofman Paul...... VB.4 Dome Balazs...... IVA.6, P4.07 Formisano Debora...... P2.34 Griggs Kim...... IA.5, P5.14 Hofmann Hans-Stefan...... P4.17 Donaldson Ken...... VA2.2 Forshed Jenny...... P3.09 Grinstaff Mark W...... VIC2.4 Holden Sophia F.D...... P2.32 Donington J S...... P5.04 Foulet Rogé Armelle...... IVC.6 Grossi Francesco...... IB.6 Holland Jimmie...... P2.30 Doolin Elizabeth...... IVB.4 Francis Roslyn...... P1.17 Grosso Federica...... P1.15, P4.26 Hollebecque Antoine...... IA.4, P1.13 Dooms Christophe...... VC.6 Franklin Peter...... IIIC.2 Grosso Stefano...... P5.05, P5.16 Honda Miki...... P3.12, P4.09, P4.16 Doyle Leona...... P1.25 Frauenfelder Thomas...... VC.2 Groussard Odile...... IVC.6 Hong Julie...... VIC2.1 Dragonieri Silvano...... P5.25 French Benjamin...... P1.19, P1.20, P2.01 Gruetzner Uwe...... P3.16, P3.17, P3.19 Hoogsteden H.C...... IIC.2, IIC.7, Duell Thomas...... P3.17 Frendl Gyorgy...... P2.04, P2.05, Grusch Michael...... IVA.6 ...... P1.30, P1.32, P3.26 Duhamel Jean-Paul...... IB.5 ...... P2.21, P3.20, P4.19 Guarrera Simonetta...... P4.21 Horion Julie...... P4.33 Duhig Edwina...... P1.35, P2.14, P4.10 Friard Sylvie...... IB.5 Guaschino Roberto...... P4.21 Hosone Masaru...... P2.26, P2.28 Dumane Vishruta...... P3.30 Fridlender Zvi G...... P1.06 Guegnon Fabien...... P4.06 Hosono Osamu...... P4.01 Dymarkowski Steven...... VC.6 Friedberg Joseph...... IIIA.4, P1.19, P1.20, Guillerme Jean-Baptiste...... P3.25 Hou Dongmei...... P1.09 ...... P2.01, P2.02, P2.03, P3.15 Howard Tim...... VB.3 Friedman Lori S...... IVA.5 Hubert Pascale...... P4.28, P4.33 E Friess Martina...... P3.02, P4.15, VC.2 H Hudson Amanda...... P1.31, P4.34 Frischknecht Lukas...... P3.02 Hughes Brett...... IVB.4 Easty David...... IC.3 Frost Felicity A...... IVC.4 Haas Andrew R...... IVB.6, P3.15 Hughes Victoria...... P1.26 Eberlein Michael...... IIB.2, IIIA.3, P3.03 Fujii Makiko...... IVA.4 Hackshaw Allan...... IVB.5 Hunt Patricia...... P2.31 Edelman J. J.B...... IIA.2, P5.14, Fujii Masanori...... P4.01 Hagemann Thorsten...... P5.13 Hunziker Roger...... VC.2 ...... WSXI.4, WSXI.5 Fujii Yoshitaka...... P2.09 Hahn Stephen M...... IIIA.4, P1.19, P1.20, Husain Aliya N...... IC.2, IVA.5 Edgar K A...... IVA.5 Fujimori Yoshihiro...... P3.07 ...... P2.01, P2.02, P2.03, P3.15 Hutanu Ionut...... P5.34 Edwards John...... P1.26, P3.22, VC.4 Fujimoto Nobukazu...... P4.01, P4.30, P5.20 Hall Robert...... IB.3 Hysi Ilir...... VB.4 Eguchi Kenji...... P5.27 Fujino Michio...... IVC.3 Hamilton Chris...... VC.7, VIB1.1 Eichhorn Martin E...... P3.16, P3.17, P3.19 Fukuda Shunichi...... P1.05, P2.08 Hamilton Peter...... P1.26 Eisen Hannah...... IIB.4, IVC.5 Fukui Takayuki...... P2.19 Hanagiri Takeshi...... P3.11 Eldem Barkin...... P4.12 Fukuoka Junya...... IVC.2 Haneda Hiroshi...... P2.09 Ellis Stephen...... IVB.5 Fukuoka Kazuya...... P1.05, P2.08, P3.07,

iMig2012.org • Abstract Book 161 I Khodayari Nazli...... P5.11 Lebaili Nemcha...... P5.15 Martinez Pablo...... P4.03, P4.04 Khong Andrea...... P2.15, P3.24 Lee Chunman...... VIA.6 Martinez-Marti Alex...... P4.03, P4.04 Inai Kouki...... IVC.2, P2.18, P4.01, Kindler H.L...... IB.2, IB.3, IC.2, IVB.3, Lee Hyun-Kyung...... IIIB.3 Martinovich Kelly...... VIC2.3 ...... P4.30, P5.20 ...... IVA.5, IVB.2, P1.16, P2.11, VC.5 Lee Lukas J...... P2.27, P5.24 Masachika Eriko...... P4.09, P4.16 Inase Naohiko...... P2.18 Kirschner Michaela B...... IC.5, IIA.2, P4.24, . Lee Sze Ting...... VC.7, VIB1.1 Mascaux Céline...... P4.28 Inchauste Suzanne...... VIC2.1 ...... P5.14, WSXI.4, WSXI.5 Lee Y.C G...... IC.4, WSXI.2 Massard Christophe...... IA.4, P1.13 Ishiguro Futoshi...... IIA.6, P2.19 Kishimoto Takumi...... P4.01, P4.30, P5.20 Lehtiö Janne...... P3.09 Massaro Tommaso...... P5.25 Ishikawa Yoshinori...... P2.19 Klabatsa Astero...... IIIB.4, P2.20, VIA.1 Leighl Natasha...... IIB.5, P3.10 Matar Kevin...... P4.10 Issaka Adamu...... P4.12 Klebe Sonja...... IA.5, IIA.2, P5.14, Lemoine Nicholas...... IVB.5 Matsumoto Seiji...... P1.05, P2.08, P3.11 ...... WSXI.4, WSXI.5 Ito Tatsuo...... P5.07 Lena Herve...... IB.5 Matsumoto Shinji...... IVC.3, P2.29 Klepetko Walter...... IVA.6, P4.07 Leon Justine...... WSXI.1 Matullo Giuseppe...... P4.21 Klikovits Thomas...... IVA.6, P4.07 Leonardi Fabio...... IIIA.5 Mazieres Julien...... IB.5

Kobayashi Hideyuki...... P2.07 index author J Leotta Attilio...... IIIC.3 Mccarthy Fiona...... P5.13 Kobayashi Hiroshi...... VIA.2 Lester Jason...... P4.29 Mccaughan Brian C...... IA.5, IIA.2, P5.14, Koch Ina...... P3.19 Jablons David...... IVA.2 Lesterhuis W. J...... IIC.5 WSXI.4, WSXI.5 Koczywas Marianna...... IB.3 Jacobs-Small Mona...... IVB.3, P3.15 Lettino Antonio...... P5.25 Mccaul Elizabeth...... P4.10 Kolev Vihren N...... P1.14 Jacobson Blake A...... IVB.7 Levin Stephen...... IA.2 Mccoy Melanie J...... P4.08 Kondo Masashi...... IIA.6 Jahan T...... IB.2 Levine Edward...... P5.30 Mcdonnell Alison M...... IB.4, IIC.4, P4.08 Kondo Nobuyuki...... P1.05, P2.08, P3.11 Jakobsen Jan Nyrop...... P4.31 Levy Brynn...... VA1.4 Mclane Amanda...... P3.28, VIB1.2 Korus Gary...... P2.03 Janes Sam...... P1.26 Li Fengzhi...... VIA.7 Mcmillan Robert...... IIA.3, IIA.4 Kozono David E...... IIIA.2, P1.08 Jantz Michael A...... P5.11 Li Hui...... IVA.2 Mechalakos James...... P3.28 Kratzke Robert A...... IVB.7, WSXI.3 Jaurand Marie-Claude...... P5.10, VB.4 Li Ming...... P1.36 Meerang Mayura...... P1.33, P3.02 Krayenbuhl Jerome...... P3.27 Jean Didier...... P5.10, VB.4 Li Quanhai...... IIIB.5, VIA.2 Mehan Michael...... IA.2 Krek Azra...... IIA.3 Jennings Cormac J...... IIA.7, P5.18 Li Xinmin...... VIC2.1 Mehta Vivek...... P3.27 Kremmidiotis Gabriel...... IVB.4 Jin Hung Chuan...... IIA.2 Libener Roberta...... P4.21 Melaiu Ombretta...... P4.22 Kristensen Claus...... P3.21 Jithesh Puthen...... P1.12 Lievense Lysanne A...... P1.32, P3.26 Melamed Jonathan...... P1.24 Krokan Hans E...... P4.23 Johansson Henrik...... P3.09 Ligeza-Poisson Catherine...... IB.5 Melis Massimo...... IIIC.3 Krug Lee M...... IB.3, IB.7, IIA.4, Judy Brendan...... P1.06 Lim Chuan Bian...... IVA.3 Mencoboni Manlio...... P1.15 ...... IVB.2, P2.30, VIB1.2 June Carl...... IVB.6 Lim Eric...... P3.22 Menegozzo Simona...... IIIC.3 Kubo Shuji...... P1.11 Lind Michael J...... IVB.5, P1.01, P5.01 Menges Craig...... P1.14 Kudelin Natalie...... IIB.2 Lindner Michael...... P3.16, P3.17, P3.19 Mensi Carolina...... IIIC.3 Kuhn Peter...... P4.02 K Ling Xiang...... VIA.7 Merler Enzo...... IIIC.3, IIIC.5, P5.26 Kumar Pavithra...... P5.05 Linton Anthony...... IA.5 Merritt Robert...... WSXI.3 Kuo Li C...... P3.28 Kaiser Dirk...... P3.05 Liu Bin...... VIA.4 Metintas Muzaffer...... P1.03, P3.09, Kuribayashi Kozo...... P3.12 Kalos Michael...... IVB.6 Liu Geoffrey...... P4.11 ...... P3.13, P4.12 Kurihara Yasuyuki...... P2.18 Kalyanaraman Balaraman...... P5.09 Liu Rong...... VIC2.4 Mick Rosemarie...... IIIA.4 Kuroda Ayumi...... P1.05, P2.08 Kamikonya Norihiko...... P1.05 Liu Xiaoxia...... P2.04, P2.05 Miettinen Markku...... VIC2.1 Kusamura Shigeki...... P5.34, VA1.2 Kamiya Hitomi...... P3.12, P4.09, P4.16 Loft Annika...... P3.21 Mikami Koji...... P3.12, P4.09, P4.16 Kusumoto Masahiko...... P5.27 Kan Shumei...... P4.18 Lucchi Marco...... P4.22 Miller Jill M...... IC.7, P5.08, VIA.5 Kwiatkowski David...... IVB.2 Kaneda Yasufumi...... VIA.6 Lui Natalie...... IVA.2 Millward Michael J...... IB.4, IIC.4, IVB.4 Kaneko Masahiro...... P5.27 Luong Fiona...... IVB.5, P5.13 Mineo Tommaso C...... P3.23

Kanemura Shingo...... P3.12, P4.16 Luttgen Madelyn...... P4.02 Mirabelli Dario...... IIIC.3, P4.21 international mesothelioma interest group L Kanteti Rajani P...... IC.2, IVA.5, P2.11 Mita Alain...... IB.3 Kao Steven C...... IA.5, P3.08, WSXI.4 Mitani Aya...... P3.20 La Spina Chiara M...... P4.27 Kapoor Veena...... P1.06 M Mitchell Paul...... IVB.4 Labby Zacariah E...... P1.16, VC.5 Kara Volkan...... P3.13, P4.12 Miura Hakutaro...... P2.18 Lackner Mark...... IVA.5 Karabela Sophia P...... IC.4 Macfarlane Marion...... P5.05, P5.16, VA2.2 Mizoi Miho...... P1.34 Ladanyi Marc...... IIA.3, IIA.4, P5.07 Karanian Marie...... IVC.2, P1.27 Macpherson Maximilian B.... IC.7, VIA.5, P5.08 Mizuguchi K. Annette...... P3.20, P4.19 Lai Chi...... IIB.6, P3.14 Kasahara Noriyuki...... P1.11 Macura Sherrill L...... VIA.3, VIA.5 Mizuno Tetsuya...... P2.19 Lake Richard...... IB.4, IIC.4, IIC.5, Katayama Hironori...... P2.26, P2.28 Moch Holger...... P3.02 ��P3.24, P4.08, VB.2, VIC2.3 Maeda Masahiro...... P1.34 Kato Katsuya...... P2.18, P4.30, P5.20 Mohamad Fahmi A...... P2.15 Lakhiani Daphne...... P3.24 Maeda Risa...... P3.12, P4.09, P4.16 Kauppinen Risto A...... VIA.3 Mohammed Kamal A...... P5.11 Lambers M. E.H...... IIC.2, IIC.7, Maeda Shotaro...... P2.26, P2.28 Molinier Oliver...... IB.5 Kawabe Takumi...... IB.3 ...... P1.30, P1.32, P3.26 Magnani Corrado...... P4.21, P5.26 Molloy Mark P...... P3.08 Kawada Ichiro...... IC.2 Lambert Didier...... P4.33 Mahaweni N.M...... P1.32 Monnet Isabelle...... IB.5 Kawaguchi Koji...... P2.19 Lambiase Antonio...... P1.10, P1.15, P2.16 Mahawi N...... IIC.7 Montero Maria Angeles...... P4.03, P4.04 Kawamata Osamu...... P2.06, P2.07, P4.13 Landau David...... P1.23, P3.18 Mahe Marc A...... P3.27, P3.29 Moon Edmund...... IVB.6 Kawamura Kiyoko...... IIIB.5, VIA.2 Landi Stefano...... P4.22 Maher John...... VIA.1 Moon Elizabeth...... P1.31 Kay E.W...... P5.18 Landry Christopher C...... VIA.3 Maintz David...... P2.22 Moore Sally...... P2.31 Kazan Steven...... P1.29, P5.21, P5.22, P5.23 Lang-Lazdunski Loic...... P1.23, P2.20, Mak Raymond H...... IIIA.2, P1.08 Kazan-Allen Laurie...... P5.33 ...... P3.18, P4.14 Maltzman J D...... IB.2 Morero Jose L...... IB.3 Keck Michaela K...... IVA.5 Langer Corey...... P3.15 Manegold Christian...... IB.1 Morimoto Chikao...... P4.01 Keegan Mitchell...... P1.14 Lansley Sally M...... IC.4 Manente Arcangela Gabriella...... P1.12, P2.12, Morin Franck...... IB.5 Kelsey Christopher...... P3.27 Lassalle Philippe...... P5.15 ...... P2.13 Moriyama Satoru...... P2.09 Kennedy Catherine...... VC.4 Laszlo Viktoria...... IVA.6 Marciniak Stefan...... P1.26 Moro Laura...... IIA.2, IIA.7, P1.12, ...... P2.12, P2.13 Kerr Keith...... P1.26 Lauchle Jennifer O...... IVB.2 Marcoux J P...... IVB.2 Moro-Sibilot Denis...... IB.5 Keshavjee Shaf...... P3.10 Lavranos Tina...... IVB.4 Margerit Sabine...... IB.1 Morrison Leanne...... P1.35, P2.14 Kestenholz Peter...... P4.15, VC.2 Lawlor Marita...... VIB1.1 Margery Jacques...... IA.4, IB.5, P1.13 Mossman Brooke T...... IC.7, P5.08, Key R. G...... P2.30 Le Pimpec-Barthes Françoise...... P5.10, VB.4 Marinaccio Alessandro...... IIIC.3 ...... P5.09, VIA.3, VIA.5 Khatri Purvesh...... WSXI.3 Le Stang Nolwenn...... IVC.2, IVC.6, P1.27 Marrinucci Dena...... P4.02 Mucchino Claudio...... IIIA.5 Khattri Arun...... IVA.5 Le Teuff Gwénaël...... IA.4, P1.13 Martin Jennifer...... P2.14 Mudrick Karen...... P3.15

iMig2012.org • Abstract Book 162 Mundt Filip...... IC.6, P1.28, P3.09, P5.17 Olsen Nola...... IIIC.2 Q Salgia Ravi...... IC.2, IVA.5, P2.11 Murer B...... P5.18 Opitz Isabelle...... IC.3, IIA.5, IIB.5, Salmons Joanne...... IIC.5 Murphy Fiona...... P5.05, P5.16, VA2.2 ...... P1.33, P3.02, P4.15, VC.2 Qadri Suhail...... P1.01 Salvini Piermario...... P4.27 Murray Jill...... IIIC.4 Orlov Sergey...... IB.3 Quispel-Janssen Josine...... VIC2.2 Sandberg Jesse...... IIB.3 Musk Arthur W...... IIIC.2, IVC.4, P2.25, Orui Hayato...... P1.05, P2.08 Santoni-Rugiu Eric...... IA.6, IIA.7, P3.01 ...... P4.05, P5.26, WSXI.1, Osada Hirotaka...... IVA.4 Santoro Armando...... P1.15, P4.26 ...... WSXI.2, WSXI.4 Ostroff Rachel M...... IA.2 R Sartorius Benn...... IIIC.4 Musti Marina...... IIIC.3, P5.25 Otsuki Taiichiro...... P3.07, P3.12, Sasai Masao...... VIA.6 Mutsaers Steven E...... IVA.3, P5.03, ...... P4.09, P4.16 Rabin Michael...... IVB.2 Sasaki Hidefumi...... P2.09 ...... WSXI.2, WSXI.4 Ottensmeier Christian...... IB.6 Ranganathan Anjana...... IVB.3 Sato Ayuko...... P2.29 Mutti Luciano...... IC.3, IIA.2, IIA.7, Ötvös Rita...... P4.32 Rao Mahadev...... VIC2.1 Sato Mitsuo...... IIA.6 ...... P1.12, P2.12, P2.13, P4.22 Ozaki Daisuke...... IVC.3 Raphael Jacques...... IA.4, P1.13, P3.29 Sauta Maria G...... P4.26, P4.27

Ozalper Hakan...... P4.12 Rapicetta Cristian...... P2.34 Sawa Yoshiki...... VIA.6 index author Ozkok Hale Basak...... P1.03, P3.13, P4.12 Rassl Doris...... P1.26 Sayan Mutlay...... P5.08, VIA.5 N O’Byrne Kenneth J...... IC.3, IIA.7 Ravn Jesper...... P3.01, P3.21 Schagen Johanna...... P2.14 O’Sullivan Brendan...... P5.01 Rea Federico...... P5.12 Schalke Berthold...... P4.17 Nabeshima Kazuki...... IVC.3, P2.29 Recio Adri...... IVB.6, P3.15 Schallenberg Gert...... IIIC.3 Nackaerts Kristiaan...... P1.18, VC.6 Reck M...... IB.2 Schedlich Lynette J...... IC.5 Nafteux Philippe...... P1.18, VC.6 P Regional Group On Schelch Karin...... IVA.6 Nagamatsu Yasuko S...... P2.33 Malignant Mesothelioma...... IIIC.5 Schenk Peter...... P4.07 Nahum Odelia...... VA1.4 Pachter Jonathan A...... P1.14 Rehauer Hubert...... IIA.5 Scherpereel Arnaud...... IB.5, P4.06, Naidoo Rishendran...... P4.10 Paci Massimiliano...... P2.34 Reid Alison...... IIIC.2, P5.26 ...... P4.28, P5.15 Naito Zenya...... P2.26, P2.28 Paciencia Maria...... IVC.2, P1.27 Reid Glen...... IA.5, IC.5, IIA.2, Schiavon Marco...... P5.12 Nakano Takashi...... P1.05, P2.08, P2.29, Padera Robert F...... VIC2.4 ...... P3.08, P4.24, P5.14, Schinwald Anja...... VA2.2 ...... P3.07, P3.11, P3.12, Padoan Marina...... P4.21 ...... WSXI.2, WSXI.4, WSXI.5 ...... P4.09, P4.16 Schirren Joachim.....IIB.2, IIIA.3, P3.03, P3.06 Pairon Jean Calude...... IVC.6 Relan Vandana...... P1.35, P2.14, P4.10, P4.24 Nakas Apostolos...... P1.07, P5.05, P5.16 Schneiter Didier...... P4.15 Pairon Jean-Claude...... P5.10, VB.4 Renier Annie...... P5.10, VB.4 Nakatani Yukio...... IVC.3 Schrump David S...... VIC2.1 Pallis Athanasios...... IB.6 Rey- Mcintyre Katrina...... IIC.3, P3.10, P4.11 Nasreen Najmunnisa...... P5.11 Schröder Heike...... P3.05 Palmer Lyle...... P4.21 Rice David...... VC.4 Natori Yuji...... P2.33 Schulz Morgan D...... VIC2.4 Parente Pereira Ana C...... VIA.1 Richards William G..... IA.7, IIB.4, IIIA.2, IVC.5, Neefjes Jacques...... VIC2.2 Schunselaar Laurel...... VIC2.2 Parienti Jean-Jacques...... IB.5 ...... P1.08, P1.21, P2.23, Neelature Sriramareddy Sathya.. P4.28, P4.33 Schwartz Ann G...... IA.3 Parnis Francis...... IVB.4 ...... VB.5, VC.3, VIC1.2 Nelson Gill...... IIIC.4 Schwartz Lawrence...... IVB.2, P2.22 Parno J...... IB.2 Ried Michael...... P4.17 Neri Monica...... P4.21 Schwed Daniel...... IVB.3 Parwani Anil V...... P1.24 Rimner Andreas...... P3.28, P3.30, VIB1.2 Neu Reiner...... P4.17 Scorsetti Marta...... P3.27 Pascovici Dana...... P3.08 Rintoul Robert C...... P1.26, P3.22 Newick Kheng...... P5.09 Scott Andrew M...... VC.7, VIB1.1 Pascucci Cristiana...... IIIC.3 Ripa Cristina...... P4.27 Nguyen Jean-Michel...... P4.06 Sebastian Lucille...... IVB.4 Pasello Giulia...... P4.26, P5.12 Riviere Alain...... IB.5 Nicastri Annalisa...... P3.04 Segal Amanda...... IVC.4, P4.05 Pass Harvey...... IA.2, IA.3, IC.3, IC.7, Robinet Gilles...... IB.5 Nieva Jorge...... P4.02 Seiwert Tanguy Y...... IVA.5, IVB.2 ...... P1.24, P5.04, P5.09, VC.4 Robinson Bruce W.S...... IB.4, IIC.4, IIC.5, Nilsson Stefan...... P1.12, P2.12 Seki Nobuhiko...... P5.27 Passmore Linda...... P4.10 ...... IVC.4, P2.15, P2.25, Nishizaki Tomoyuki...... P4.09 ...... P3.24, P4.05, WSXI.1, Sekido Yoshitaka...... IVA.4 Pastan Ira...... IB.2 Niwa Hiroshi...... P2.09 ...... WSXI.2, WSXI.4 Sekine Ikuo...... VIA.2

Patel Manish R...... IVB.7 international mesothelioma interest group Nixon Lisette...... P4.29 Robinson Cleo...... VB.2, VIC2.3 Selch Michael...... IIB.6, P3.14 Patsios Demetris...... P4.11 Nogi Yoshitaka...... P3.12, P4.09, P4.16 Rodriquez Cristina...... IB.3 Sellitri Francesco...... P3.23 Pavlakis Nick...... IVB.4, P1.31, P3.08, P4.34 Nolan Luke...... IVB.5 Rolli Luigi...... IIIA.5 Sensakovic William F...... VC.5 Pei Jianming...... VA1.4 Nonaka Daisuke...... IC.3 Romanelli Antonio...... IIIC.3 Sensi Elisa...... P4.22 Peikert Tobias...... IVB.7 Noraian Cynthia...... P1.22 Rorie Anne...... IIB.3, IIB.6, P3.14 Serke Monika...... P3.05 Pel Marcella...... WSXI.5 Nowak Anna K...... IB.4, IIC.4, IIC.5, Rosa Fabio...... P4.21 Setinek Ulrike...... IVA.6 Pentakota Sujatha...... P2.21 ...... IVB.4, IVC.4, P1.16, Rosenzweig Kenneth E... P3.28, P3.30, VIB1.2 Sgarbi Giorgio...... P2.34 Perigaud Christian...... P3.29, P4.06 ...... P1.17, P4.08, VIC2.3 Ross Helen...... IB.3 Shah Riyaz...... P3.22 Perkins Timothy N...... IC.7, VIA.5 Rossi Leonardo...... P1.12 Shapiro Irina M...... P1.14 Persigehl Thorsten...... P2.22 Rossoni Gilda...... P1.10, P2.16 Sharma Sunil...... IB.3 Peters Lydnsey...... P4.34 O Roulois David...... P3.25 Sharpe Kevin...... P5.13 Peto Julian...... P3.22 O’Brien Carol...... IVA.5 Rousvoal Thomas...... IVC.6 Sheaff Michael...... IIIB.4, IVB.5 Phillips Melissa...... IVB.5, P5.13 O’Brien Mary...... IB.6 Ruckdeschel John...... IB.3 Shi Yangdong...... IIA.5 Piccolini Ezio...... P4.21 O’Byrne Kenneth...... P1.12 Rudd Robin...... IVB.5 Shibata Eisuke...... P3.12, P4.16 Picquenot Jean Michel...... IVC.6 O’Donnell Dearbhaile...... IC.3 Ruffini Enrico...... P4.21, VC.4 Shigematsu Yoshiki...... P3.11 Pinton Giulia...... P1.12, P2.12, P2.13 O’Donnell Eimear...... IIA.7 Ruiz William F...... P5.21 Shimada Hideaki...... IIIB.5, IVC.3, VIA.2 Pirker Christine...... IVA.6 O’Grady A...... P5.18 Rusca Michele...... IIIA.5 Shingyoji Masato...... IIIB.5 Piro Roberto...... P2.34 O’Shannessy Dan...... IB.2 Rusch Valerie...... IIA.4, P2.30, VC.4, VIB1.2 Shitara Masayuki...... P2.09 Planchard David...... IA.4, P1.13 O’Sullivan Brenda...... P4.11 Russell Dale...... P2.31 Shouldis Jennifer...... IVB.2, IVB.3, VC.5 Pohl Wolfgang...... P4.07 Ohar Jill...... P5.30, VB.3 Russell Stephen J...... IVB.7 Shrager Joseph...... WSXI.3 Polo Valentina...... P4.26 Ohkuwa Hisaya...... P3.12 Rykers Kym...... VIB1.1 Shukla Arti...... IC.7, P5.08, P5.09, Popat Sanjay K...... IB.6, P3.22 Ohnuma Kei...... P4.01 Røe Oluf D...... P4.23, P5.36 ...... VIA.3, VIA.5 Okada Asuka...... P3.07, P3.12, P4.09, P4.16 Porta Camillo...... P1.15 Silic-Benussi Micol...... P5.12 Okazaki Yasumasa...... P5.06 Potzger Tobias...... P4.17 Sima Camelia S...... IB.7, IIA.4 Praet Marleen...... IVC.2 Okiror Lawrence...... P1.23, P3.18 Simone Charles B...... P1.19, P1.20, Okuda Katsuhiro...... P2.09 Predina Jarrod D...... P1.06 S ...... P2.01, P2.02, P2.03, P3.15 Okumura Yoshitomo...... P1.05, P2.08, P3.11 Prele Cecilia M...... IVA.3 Singhal Sunil...... P1.06 Okuwa Hisaya...... P4.09, P4.16 Price Allan...... IB.6 Sagan Christine...... IVC.6, P4.06 Sklarek Juergen...... P3.16, P3.17 Olevsky Olga M...... IIB.6, P3.14 Putt Mary...... P1.19, P1.20, P2.01 Sakai Fumikazu...... P2.18 Soerensen Jens B...... IA.6

iMig2012.org • Abstract Book 163 Sollis Elliot...... VB.2 Tamura Kunihiro...... P3.07, P3.12, Vansteenkiste Johan...... VC.6 ...... VB.5, VC.3, VIC1.3 Soltermann Alex...... IC.3, IIA.5, P3.02 ...... P4.09, P4.16 Varano Julius...... IC.4 Yokoi Kohei...... IIA.6, P2.19 Song Misa...... P2.29 Tan Ai Ling...... IC.4 Vardy Janette...... IA.5 Yoneda Kazue...... P1.05, P3.11 Song Xiaomin...... P3.08 Tan Yongqiang...... P2.22 Varghese Sheelu...... P5.35 Yorke Ellen...... P3.28, P3.30, VIB1.2 Sorensen Jens Benn...... P3.21, P4.31 Tanahashi Masayuki...... P2.09 Verardo Marina...... IIIC.3 Yoshida Kenya...... IIA.6 Soria Jean-Charles...... IA.4, P1.13 Tanaka Fumihiro...... P1.05, P2.08, P3.11 Verbeken Eric...... P1.18, VC.6 Yoshii Naoko...... P2.09 Sougawa Nagako...... VIA.6 Tanaka Ichidai...... IVA.4 Verschakelen Johny...... VC.6 Yoshikawa Yoshie...... P3.07 Spicer James...... IVB.5, P1.23, P3.18 Tangy Frédéric...... P3.25 Vidal Christian M...... P1.14 Yoshino Naoyuki...... P2.26 Squadroni Michela...... P4.27 Taniere Phillipe...... P5.01 Vignaud Jean Michel...... IVC.2, IVC.6 Young Ann M...... IVB.2 Sriram Krishna B...... P4.10 Taniguchi Tetsuo...... P2.19 Vigneswaran Wickii...... IC.2 Yukiue Haruhiro...... P2.09 Stahel Rolf...... IIA.5, P1.33, P3.04 Tanno Masataka...... P2.26, P2.28 Vitek Olga...... P3.04 Yumuk Fulden...... P1.03, P3.13, P4.12 Stahlhut Leandra...... P2.22 Tao Sha...... VB.3 Vivian Justin...... P2.15 Yun Zhihong...... IIC.3, IIC.6, P1.36,

Stallmann Sonja...... P3.03, P3.05, P3.06 Tatsumi Koichiro...... IIIB.5, VIA.2 Voglino Floriana...... P4.21 ...... P3.10, P4.11 index author Starr Tatiana...... P2.30 Taub Robert N...... VA1.4 Yusa Toshikazu...... IVC.3 Stathopoulos Georgios T...... IC.4 Taylor Henry...... P3.18 W Steele Jeremy P...... IVB.5, P3.18, Taylor Paul...... IVB.5 ...... P3.22, P5.13 Tenconi Sara...... P2.34 Wagner Andrew...... IVB.2 Z Steinbacher Jeremy L...... VIA.3 Terada Takayuki...... P3.12, P4.09, P4.16 Waikar Sushrut...... P3.20, P4.19 Sterman Daniel...... IIIA.4, IVB.6, P1.19, Testa Joseph...... P1.14, VA1.4 Wakelee Heather...... WSXI.3 Zalcman Gerard...... IB.5 ...... P1.20, P2.01, P2.02, Tewaternaude Jim...... IIIC.4 Wallace W. Dean...... IIB.6, P3.14 Zanichelli Stefano...... IIIA.5 ...... P2.03, P3.15 Thayaparan Thivyan...... VIA.1 Waller David A...... P1.07, P3.22, VC.4 Zauderer Marjorie G...... IB.7, IIA.4, IVB.2, Stern Eric...... P4.33 Thies Svenja...... IIA.5, P3.02 Wallin B...... IB.2 ...... P2.30, VIB1.2 Sterrett Greg F...... IVC.4 Thivolet Bejui Francoise...... IVC.6 Wallin Jeffrey J...... IVA.5 Zefkili Sophia...... P3.29 Sterzing Florian...... P3.27 Thomas Warren...... IIA.7, P5.18 Walsh Amy...... VIC2.3 Zhang Mary...... VIC2.1 Stevenson James...... IIIA.4, IVB.3, P1.19, Thompson Joyce K...... VIA.5 Wang Eric J...... P1.19, P1.20, P2.01, Zhang Siyu C...... IA.3 ...... P1.20, P2.01, P2.02, Thompson Philip J...... IVA.3, P5.03 ...... P2.02, P2.03 Zhang Xinbo...... IA.3 ...... P2.03, P3.15 Tonoli Sandro...... P3.27 Wang Jung-Der...... P5.24 Zhao Binsheng...... P2.22 Stewart Alex...... IA.2 Tookman Laura...... P5.13 Wang Yaoyu E...... IA.7, VB.5, VIC1.2, Zhao Yidan...... IIC.6, P1.36 Stinco Sergio...... P4.26, P4.27 Torii Ikuko...... P1.05, P2.29 ...... VIC1.3 Zhibin Gao...... P5.36 Stockler Martin...... IVB.4 Torrejon Davis...... P4.03, P4.04 Weder Walter...... IIA.5, P1.33, P3.02, Ziegler Annemarie...... P3.04 Straus Christopher...... VC.5 Toyokuni Shinya...... P5.06 ...... P4.15, VC.2, VC.4 Zielinski Christoph...... P4.07 Stuart O. Anthony...... VA1.3 Trafficante Luana...... IIIC.3 Weir Chris...... P1.31, P4.34 Zimilover Adam...... VA1.4 Su Yang...... VIA.4 Treasure Tom...... P3.22 Whelan Nancy...... P1.24 Zimling Zarah G...... IA.6, IIA.7 Sugarbaker David J...... IA.7, IIB.4, IIIA.2, Wieczorek Andrezej...... P1.01 Zong Fang...... IC.6, P5.17 ...... IIIB.2, IIIB.3, IVC.5, P1.08, Tretiakova Maria...... IC.2 Willems Luc...... P4.28, P4.33 ...... P1.21, P2.04, P2.05, P2.21, Tsao Ming...... P1.36, P4.11 Zsiklavari Zsolt...... P4.17 ...... P2.23, P2.24, P3.20, P4.19, Tseng Katty...... IVA.2 Williams Marissa...... WSXI.5 Zucali Paolo...... P1.15, P4.26 ...... P4.20, VC.3, VIC1.2, VIC1.3 Tsicopoulos Anne...... P5.15 Williams Stephen...... IA.2 Zucchi Luigi...... P2.34 Sugarbaker Paul H...... VA1.3 Tsubota Noriaki...... P1.05, P2.08, P4.16 Willis Anne E...... P5.05, P5.16, VA2.2 Zucman-Rossi Jessica...... P5.10, VB.4 Sugarbaker Peter E...... VB.5 Tsujimura Tohru...... P1.05, P2.29 Windsor Morgan...... P4.10 Zuo Zhixiang...... IVA.5 Suh Robert...... IIB.3, IIB.6 Tumino Rosario...... IIIC.3 Winfrey Olivia...... IIB.4, IIIA.2, IVC.5 Sun Xiao Ming...... P5.05, P5.16, VA2.2 Tunariu Nina...... IVB.2 Winter Maximilian P...... P4.07

Supko Jeffry...... IIB.4 Wolf Andrea S...... IVC.5, P1.08, P1.21, VC.3 international mesothelioma interest group Suzuki Eriko...... P2.09 Wollscheid Bernd...... P3.04 Sylvestre Alma...... P3.29 U Woo Samantha...... VIC2.3 Szatmári Tünde...... IC.6, P5.17 Wouters Johan...... P4.33 Szekely Laszlo...... P4.32 Ubiratan Moura...... IIA.5 Wozniak Antoinette...... IB.3 Szlosarek Peter...... IVB.5, P1.26, P5.13 Ueda Mamoru...... P4.18 Wright Casey M...... P4.24, WSXI.5 Szulkin Adam...... P4.32 Ugolini Donatella...... P4.21 Wu Abraham J...... P3.28, P3.30, VIB1.2 Szyszko Teresa...... IVB.5 Upham Trevor...... VIC2.1 Wu Di...... IVC.3 Sørensen Jens Benn...... IIA.7, P3.01 Uramoto Hidetaka...... P3.11 Wu Licun...... IIC.3, IIC.6, P1.36, �� P3.10, P4.11 Urso Loredana...... P5.12 Wu Mu-Zon...... P2.27 Usami Noriyasu...... IIA.6 T Wylie Ben...... P3.24 Xi Sichuan...... VIC2.1 Xu Jianfeng...... VB.3 Taatjes Douglas J...... VIA.3 V Tabata Chiharu...... P3.07, P3.12, Xu Qunli...... P1.14 Xu Wei...... P4.11 ...... P4.09, P4.16 Vacca Anna Rosa...... P1.12 Xu Yue...... WSXI.3 Tabot Denis C...... IB.6 Valenti Daniela...... P1.12 Tacconi Federico...... P3.23 Vallely Michael P...... IIA.2, P5.14, Tada Yuji...... IIIB.5, IVC.3, VIA.2 ...... WSXI.4, WSXI.5 Tagawa Kohei...... P4.18 Van Der Most Robbert G...... P4.08 Y Tagawa Masatoshi...... IIIB.5, IVC.3, VIA.2 Van Der Stagen Sjoukje...... VIA.1 Tagawa Tetsuzo...... IIC.3, P3.10, Van Meerbeeck Jan P...... IB.1, IB.6, Yaguchi Takahiro...... P4.09 ...... P4.11, P4.15 ...... P4.14, VC.4 Yamada Shusai...... P3.07, P3.12, Takagi-Kimura Misato...... P1.11 Van Raemdonck Dirk...... VC.6 ...... P4.09, P4.16 Takenaka Masaru...... P3.11 Van Schalkwyk May...... VIA.1 Yamada Taketo...... P4.01 Takeshima Yukio...... P2.18, P4.01, Van Zandwijk Nico...... IA.5, IC.5, IIA.2, Yamaguchi Takuhiro...... P5.27 ...... P4.30, P5.20 ...... P3.08, P4.24, P5.14, Yamashita Ryo...... IIA.6 Takiguchi Yuichi...... IIIB.5, VIA.2 ...... WSXI.2, WSXI.4, WSXI.5 Yang Ian A...... P1.35, P2.14, P4.10 Takuwa Teruhisa...... P1.05, P2.08, P3.11 Vanderlaan Paul...... P2.24 Yano Motoki...... P2.09 Tamamoto Atsuko...... P1.11 Vandermeers Fabian...... P4.28, P4.33 Yeap Beow Y...... IA.7, P1.21, P4.20,

iMig2012.org • Abstract Book 164